Sweet Basil (Ocimum basilicum L.)―A Review of Its Botany, Phytochemistry, Pharmacological Activities, and Biotechnological Development

Abstract

An urgent demand for natural compound alternatives to conventional medications has arisen due to global health challenges, such as drug resistance and the adverse effects associated with synthetic drugs. Plant extracts are considered an alternative due to their favorable safety profiles and potential for reducing side effects. Sweet basil (Ocimum basilicum L.) is a valuable plant resource and a potential candidate for the development of pharmaceutical medications. A single pure compound or a combination of compounds exhibits exceptional medicinal properties, including antiviral activity against both DNA and RNA viruses, antibacterial effects against both Gram-positive and Gram-negative bacteria, antifungal properties, antioxidant activity, antidiabetic potential, neuroprotective qualities, and anticancer properties. The plant contains various phytochemical constituents, which mostly consist of linalool, eucalyptol, estragole, and eugenol. For centuries, community and traditional healers across the globe have employed O. basilicum L. to treat a wide range of ailments, including flu, fever, colds, as well as issues pertaining to digestion, reproduction, and respiration. In addition, the current research presented underscores the significant potential of O. basilicum-related nanotechnology applications in addressing diverse challenges and advancing numerous fields. This promising avenue of exploration holds great potential for future scientific and technological advancements, promising improved utilization of medicinal products derived from O. basilicum L.

Full text

Citation: Azizah, N.S.; Irawan, B.;
Kusmoro, J.; Safriansyah, W.; Farabi,
K.; Oktavia, D.; Doni, F.; Miranti, M.
Sweet Basil (Ocimum basilicum L.)—A
Review of Its Botany, Phytochemistry,
Pharmacological Activities, and
Biotechnological Development.
Plants 2023,12, 4148. https://
doi.org/10.3390/plants12244148
Academic Editors: Dušanka Kiti´c,
Katarina Šavikin and
Milica Randjelovi´c
Received: 3 November 2023
Revised: 9 December 2023
Accepted: 11 December 2023
Published: 13 December 2023
Copyright: © 2023 by the authors.
Licensee MDPI, Basel, Switzerland.
This article is an open access article
distributed under the terms and
conditions of the Creative Commons
Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
4.0/).
plants
Review
Sweet Basil (Ocimum basilicum L.)—A Review of Its Botany,
Phytochemistry, Pharmacological Activities, and
Biotechnological Development
Nabilah Sekar Azizah 1, Budi Irawan 1, Joko Kusmoro 1, Wahyu Safriansyah 2, Kindi Farabi 2, Dina Oktavia 3,
Febri Doni 1and Mia Miranti 1, *
1Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran,
Jatinangor 45363, Indonesia; nabilah17004@mail.unpad.ac.id (N.S.A.); budi.irawan@unpad.ac.id (B.I.);
joko.kusmoro@unpad.ac.id (J.K.); febri@unpad.ac.id (F.D.)
2Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran,
Jatinangor 45363, Indonesia; wahyu17002@mail.unpad.ac.id (W.S.); kindi.farabi@unpad.ac.id (K.F.)
3Department of Transdisciplinary, Graduate School, Universitas Padjadjaran, Bandung 40132, Indonesia;
dina.oktavia@unpad.ac.id
*Correspondence: mia.miranti.rustama@unpad.ac.id
Abstract:
An urgent demand for natural compound alternatives to conventional medications has
arisen due to global health challenges, such as drug resistance and the adverse effects associated
with synthetic drugs. Plant extracts are considered an alternative due to their favorable safety
profiles and potential for reducing side effects. Sweet basil (Ocimum basilicum L.) is a valuable
plant resource and a potential candidate for the development of pharmaceutical medications. A
single pure compound or a combination of compounds exhibits exceptional medicinal properties,
including antiviral activity against both DNA and RNA viruses, antibacterial effects against both
Gram-positive and Gram-negative bacteria, antifungal properties, antioxidant activity, antidiabetic
potential, neuroprotective qualities, and anticancer properties. The plant contains various phytochem-
ical constituents, which mostly consist of linalool, eucalyptol, estragole, and eugenol. For centuries,
community and traditional healers across the globe have employed O. basilicum L. to treat a wide
range of ailments, including flu, fever, colds, as well as issues pertaining to digestion, reproduction,
and respiration. In addition, the current research presented underscores the significant potential of
O. basilicum-related nanotechnology applications in addressing diverse challenges and advancing
numerous fields. This promising avenue of exploration holds great potential for future scientific and
technological advancements, promising improved utilization of medicinal products derived from
O. basilicum L.
Keywords: Ocimum basilicum L.; antiviral; antifungal; anticancer; nanotechnology; biotechnology
1. Introduction
In recent years, natural plant-based products have emerged as a valuable global
resource for the development and innovation of novel drugs [
1
,
2
]. Hence, exploring bioac-
tive compounds from various sources, including plants, might be an excellent method
for discovering new potential drugs [
3
]. This is because the current availability of raw
materials for drug discovery and development, pharmacophores, and a framework for
effective medications for a wide range of clinical indications is notably limited [
4
]. Hence,
ethnopharmacological studies are of great significance, as they harness traditional knowl-
edge to effectively screen and improve the chances of discovering novel drugs [5].
Basil (Ocimum basilicum L.) is one of the species in the Lamiaceae family, which is well
known for having a wide variety of medicinal properties [
6
]. The plant is traditionally
recognized for its utilization for both culinary and perfumery purposes [
7
]. For example,
Plants 2023,12, 4148. https://doi.org/10.3390/plants12244148 https://www.mdpi.com/journal/plants

Plants 2023,12, 4148 2 of 25
in the province of East Nusa Tenggara, Indonesia, the Tetun people frequently consume
fresh, raw O. basilicum L. leaves in order to treat malaria [
8
]. In addition, it is also used
for treating rheumatism, high cholesterol, hypertension, headaches, and stroke in the
Indonesian province of North Sumatra by the Batak Karo people [
9
]. O. basilicum L. leaves
also find application as an anti-helminthic remedy among the Muna Tribe in the province
of Southeast Sulawesi, Indonesia [10].
Some of the uses stated are associated with the main constituents found in O. basilicum
L. plant parts, which include linalool, eugenol, geranial, methyl eugenol, 1,8-cineole,
and other compounds [
11
]. These compounds were found to play important roles as
antimicrobials, antioxidants, anticancer agents, and antidiabetics [
12
]. Certain chemical
compounds, specifically linalool and eugenol, are in great demand at present. This urgency
arises from the resistance exhibited by Staphylococcus aureus, which is known for its ability
to create biofilms [13].
This article aims to present a comprehensive overview of the current and ongoing
progress in the use of O. basilicum L. for medical purposes in human and animal healthcare,
with the aim of serving as a guide, which traces the historical uses of O. basilicum L. from
ethnopharmacology to biotechnological development. Additionally, this article aims to pro-
mote further clinical research efforts and the development of pharmaceutical formulations
using O. basilicum L. as a valuable resource.
The investigation commences by examining the morphological and chemical compo-
sitions of O. basilicum L. Subsequently, we proceed to gather empirical evidence derived
from ethnomedicinal data originating from diverse regions and continents around the
world. Moreover, there are substantial data supporting the therapeutic benefits of this plant
species from the perspectives of microbiology and biomedicine. Finally, we will explore the
future prospects of nanotechnology in this field and investigate the strategies to enhance
its metabolite production.
2. Methodology
A literature survey using various keywords, such as “Ocimum basilicum L.”, “antiviral”,
“phytochemical constituents”, “ethnomedicinal use”, was conducted in scientific databases,
including Scopus, ResearchGate, and Google Scholar. Out of all the collected publications,
156 underwent thorough evaluation and included research articles, review articles, and
book chapters. From this array of scientific resources, our focus was directed toward the
antimicrobial activity, phytochemical constituents, and biotechnological advancements of
O. basilicum L.
3. Ocimum basilicum L. Ecology and Morphology
Basil is renowned for its ability to thrive in diverse temperature ranges and geographi-
cal regions, making it a globally cultivated herb [
14
]. The genus Ocimum, which belongs
to the Lamiaceae (Labiatae) family, has distribution throughout tropical and subtropical
America, Africa, and Asia continents [
15
]. Ocimum has over 150 species and is extensively
cultivated in countries such as Indonesia, India, Morocco, France, Hungary, Greece, and
Egypt [
16
]. Although it is grown as a common garden herb, basil is most likely native to
Asia and Africa. It is believed that Alexander the Great (356–323 BCE) brought it from
India to ancient Greece, to England in the middle of the 1500s, and to the United States in
the early 1600s. Many countries, including Egypt, India, Indonesia, Mexico, and the United
States, produce this plant commercially for the market [17].
Sweet basil is an annual herb with dense foliage and a variety of aromatic compo-
nents [
18
]. This plant thrives in an agroclimatic environment, with temperatures ranging
from 7 to 27
◦
C, annual precipitation from 0.6 to 4.3 m, and soil pH from 4.3 to 8.2. This plant
requires low maintenance, and it is easy to grow in indoor and outdoor settings [
17
]. Al-
though it can be damaged by frost and temperatures below freezing, this species flourishes
under conditions of long daylight with full sun and well-drained soil [19].

Plants 2023,12, 4148 3 of 25
The plant can grow up to 0.6 m in height, with lateral branches creating an angle of
more than 30
◦
with the main branch. The stem is round–quadrangular, glabrous (smooth,
hairless), or puberulent (fine short hairs), concentrated on the two opposing faces of the
stem (Figure 1A). Inflorescence is dense (Figure 1B), arranged around a point on an axis
up to 12 mm apart; the axis is pubescent and with a total of six flowers surrounding the
apex (Figure 1C). The leaves are green, the apex mostly acute or acuminate; the shape is
ovate or elliptic ovate; the size is about 15–50
×
5–25 mm; the leaf margin is entirely or
sparsely serrate and with a glandular–punctate shape. The petiole is about 20 mm long and
pubescent (covered with soft short hair) (Figure 1D). The corolla with a white or pinkish
color tube—about 7–8 mm long—is funnel-shaped (Figure 1E). The calyx pilose (covered
with soft long hair or pubescent) has a dense ring of hairs at the throat, and a fruiting
calyx is about 6 mm long. The stamen has tufted hairs near the base. The nutlets are dark
brown in color, with an elliptic shape, and they produce mucilage upon interaction with
water [20,21].
Plants 2023, 12, x FOR PEER REVIEW 3 of 26
Although it can be damaged by frost and temperatures below freezing, this species our-
ishes under conditions of long daylight with full sun and well-drained soil [19].
The plant can grow up to 0.6 m in height, with lateral branches creating an angle of
more than 30° with the main branch. The stem is round–quadrangular, glabrous (smooth,
hairless), or puberulent (ne short hairs), concentrated on the two opposing faces of the
stem (Figure 1A). Inorescence is dense (Figure 1B), arranged around a point on an axis
up to 12 mm apart; the axis is pubescent and with a total of six owers surrounding the
apex (Figure 1C). The leaves are green, the apex mostly acute or acuminate; the shape is
ovate or elliptic ovate; the size is about 15–50 × 5–25 mm; the leaf margin is entirely or
sparsely serrate and with a glandular–punctate shape. The petiole is about 20 mm long
and pubescent (covered with soft short hair) (Figure 1D). The corolla with a white or pink-
ish color tube—about 7–8 mm long—is funnel-shaped (Figure 1E). The calyx pilose (cov-
ered with soft long hair or pubescent) has a dense ring of hairs at the throat, and a fruiting
calyx is about 6 mm long. The stamen has tufted hairs near the base. The nutlets are dark
brown in color, with an elliptic shape, and they produce mucilage upon interaction with
water [20,21].
Figure 1. (A) O. basilicum L. plant habit in a plantation area of Parongpong subdistrict, Bandung,
Indonesia. (B) Full O. basilicum L. inorescence. (C) Separate photograph detailed section of ino-
rescence. (D) Leaf. (E) Flower (photographs courtesy of Nabilah Sekar Azizah).
A
B
D
C
E
Figure 1.
(
A
)O. basilicum L. plant habit in a plantation area of Parongpong subdistrict, Bandung,
Indonesia. (
B
) Full O. basilicum L. inflorescence. (
C
) Separate photograph detailed section of inflores-
cence. (D) Leaf. (E) Flower (photographs courtesy of Nabilah Sekar Azizah).
4. Phytochemical Constituents
The type of chemotype can affect the main chemical constituent of O. basilicum L. [
22
].
Varga et al. discovered five chemotypes, among which (A) linalool (
15
); (B) linalool
(
15
)/trans-
α
-bergamotene (
29
); (C) linalool (
15
)/methyl chavicol (
21
); (D) linalool (
15
)/trans-

Plants 2023,12, 4148 4 of 25
methyl cinnamate (
25
); and (E) methyl chavicol (
21
). Based on the distribution in the
regions, chemotypes A and C are European chemotypes; D is a tropical chemotype; and E
is a Reunion chemotype [
22
]. The chemical constituents of O. basilicum L. were dominated
by compounds from the phenylpropanoid and monoterpenoid classes [23].
One of the most crucial aspects determining the quality of essential oil depends on the
method by which it was adopted. In some studies, conventional extraction methods, such
as Soxhlet, hydrodistillation, steam distillation, solvent extraction, and a combination of
steam and solvent extraction, are still being used [
24
]. However, there are promising green
extraction methods, including microwave-assisted extraction (MAE), ultrasound-assisted
extraction (UAE), high-pressure-assisted extraction (HAE), supercritical and subcritical
fluid extraction, electrically assisted extractions, and enzyme-assisted extraction [
25
]. Green
extraction techniques address the challenges by offering several advantages, including
cutting down on the use of organic solvents, ease of use due to the simplicity, time efficiency,
and cost effectiveness in the extraction process [26].
The O. basilicum L. extract and essential oil contain classes of chemical compounds,
mainly terpenoids, such as oxygenated sesquiterpenes, oxygenated monoterpenes, sesquiter-
pene hydrocarbons, monoterpene hydrocarbons, and non-terpene derivatives. Furthermore,
this plant contains phenylpropanoid compounds, including eugenol, methyl eugenol, chav-
icol, estragole, and methyl cinnamate [
27
,
28
]. Monoterpene, geraniol, myrtenol, pinene,
camphor, and borneol hold potential for medical applications [29].
The analysis of O. basilicum L. methanolic leaves’ extract, obtained by sonication,
showed the presence of various polyphenol compounds, such as caffeic acid, caftaric acid,
3,4-dihydroxyphenylacetic acid, ferulic acid, rosmarinic acid, and rutoside (rutin) [
30
].
Phenolic compounds have been widely recognized for their beneficial properties and
applications in the medical field [
31
]. Furthermore, O. basilicum L. extract was also found
to contain phytosterols, such as β-sitosterol, stigmasterol [32], and campesterol [33].
Studies on the chemical constituents of O. basilicum L. are shown in Table 1, repre-
senting its essential oil and extract with various extraction and identification methods.
Meanwhile, Figures 2and 3show the chemical structures of the chemical compounds
presented in Table 1.
Table 1. Chemical constituents from extracts and essential oil of O. basilicum L.
No. Chemical Compound Molecular
Weight Source Extraction and Identification
Method Reference
Monoterpene Hydrocarbon
1. α-pinene 136.23 g/mol Leaf
Hydrodistillation, GC-MS
Maceration 24 h, GC-MS
Hydrodistillation, solvent
extraction, GC-MS
[34]
[35]
[36]
2. β-Myrcene 136.23 g/mol Leaf Hydrodistillation, GC-MS [34]
3. Citral 153.23 g/mol Leaf Hydrodistillation, GC-MS [34]
4. Camphene 136.23 g/mol Leaf Maceration, GC-MS
SFME, hydrodistillation 1 h, GC-MS
[32]
[37]
5. Terpineol 154.25 g/mol Leaf Maceration, GC-MS [32]
6. Linalyl acetate 196.29 g/mol Leaf Maceration, GC-MS [32]
7. cis-Sabinene hydrate 154.25 g/mol Leaf Maceration, GC-MS [32]
8. (−)-trans-Pinocarvyl acetate 194.27 g/mol Leaf Maceration, GC-MS [32]
9. Limonene dioxide 168.23 g/mol Leaf Maceration, GC-MS
Hydrodistillation, GC-MS
[32]
[38]
10. Geraniol 154.25 g/mol Leaf Hydrodistillation, GC-MS [38]
11. Carvone 150.22 g/mol Leaf Hydrodistillation 3 h, GC-MS [39]
12. Myrtenol 152.23 g/mol Leaf Hydrodistillation, GC-MS [40]
13. Fenchone 152.23 g/mol Aerial parts Maceration 24 h, GC-MS
Hydrodistillation 4 h, GC-MS
[35]
[28]

Plants 2023,12, 4148 5 of 25
Table 1. Cont.
No. Chemical Compound Molecular
Weight Source Extraction and Identification
Method Reference
Oxygenated Monoterpene
14. l-Menthol 156.26 g/mol Leaf Hydrodistillation, GC-MS [34]
15. Linalool 154.25 g/mol Leaf Maceration, GC-MS
Hydrodistillation, GC-MS
[32]
[38]
16. trans-linalool oxide 170.25 g/mol Leaf Maceration, GC-MS [32]
17. cis-Linalool-oxide 213.27 g/mol Leaf Hydrodistillation, GC-MS
Maceration, GC-MS
[34]
[32]
18. Camphor 152.23 g/mol Leaf
Maceration, GC-MS
Maceration 24 h, GC-MS
SFME, hydrodistillation 1 h, GC-MS
[32]
[35]
[37]
19. Neral 152.23 g/mol Aerial parts Maceration 24 h, GC-MS
SFME, hydrodistillation 1 h, GC-MS
[35]
[37]
20. 1,8-cineole (Eucalyptol) 154.25 g/mol Leaf
Hydrodistillation, GC-MS
Maceration, GC-MS
Hydrodistillation, solvent
extraction, GC-MS
[34]
[32]
[36]
21. Estragole (Methyl chavicol) 148.20 g/mol Leaf, flower,
inflorescence
Hydrodistillation, GC-MS
Hydrodistillation 3 h, GC-MS
Maceration 24 h, GC-MS
Hydrodistillation, solvent
extraction, GC-MS
[34]
[39]
[35]
[36]
22. Eugenol 164.20 g/mol Leaf, aerial
parts
Hydrodistillation, GC-MS
Maceration 24 h, GC-MS
SFME, hydrodistillation 1 h, GC-MS
[34]
[35]
[37]
23. Methyl eugenol 173.23 g/mol Leaf Hydrodistillation 3 h, GC-MS
Maceration 24 h, GC-MS
[39]
[35]
24. Bornyl acetate 196.29 g/mol Aerial parts Hydrodistillation 4 h, GC-MS [28]
25. Methyl cinnamate 162.18 g/mol Leaf Hydrodistillation, GC-MS [40]
Sesquiterpene Hydrocarbon
26. Copaene 204.35 g/mol Leaf Hydrodistillation, GC-MS [34]
27. Neoisolongifolene 202.33 g/mol Leaf Hydrodistillation, GC-MS [34]
28. α-Bergamotene 204.35 g/mol Leaf
SFME, hydrodistillation 1 h, GC-MS
[37]
29. trans-.alpha.-Bergamotene 204.35 g/mol Leaf
Hydrodistillation, GC-MS
Hydrodistillation, solvent
extraction, GC-MS
Hydrodistillation, GC-MS
[34]
[36]
[38]
30. β-farnesene 204.35 g/mol Leaf Hydrodistillation, GC-MS [38]
31. Alloaromadendrene 204.35 g/mol Leaf Hydrodistillation, GC-MS [34]
32. γ-Cadinene 204.35 g/mol Leaf
SFME, hydrodistillation 1 h, GC-MS
[37]
33. Humulene 204.35 g/mol Leaf Hydrodistillation, GC-MS [34]
34. α-Humulene 204.35 g/mol Leaf
SFME, hydrodistillation 1 h, GC-MS
[37]
35. α-Copaene 204.35 g/mol Leaf
SFME, hydrodistillation 1 h, GC-MS
[37]
36. β-Copaene 204.35 g/mol Leaf Hydrodistillation, GC-MS [34]
37. β-Bisabolene 204.35 g/mol Leaf Hydrodistillation, GC-MS [34]
38. cis-muurola-3,5-diene 204.35 g/mol Leaf Hydrodistillation, GC-MS [34]
39. cis-.alpha.-Bisabolene 204.35 g/mol Leaf Hydrodistillation, GC-MS [34]
40. α-Cubebene 204.35 g/mol Leaf, aerial
parts
Hydrodistillation, GC-MS
Maceration 24 h, GC-MS
Hydrodistillation, solvent
extraction, GC-MS
[40]
[35]
[36]
41. Germacrene B 204.35 g/mol Leaf Hydrodistillation, GC-MS [40]
42. Germacrene D 204.35 g/mol Leaf
Hydrodistillation, GC-MS
Hydrodistillation 4 h, GC-MS
Maceration 24 h, GC-MS
Hydrodistillation, GC-MS
[40]
[28]
[35]
[38]

Plants 2023,12, 4148 6 of 25
Table 1. Cont.
No. Chemical Compound Molecular
Weight Source Extraction and Identification
Method Reference
43. β-Elemene 204.35 g/mol Leaf
SFME, hydrodistillation 1 h, GC-MS
[37]
44. β-Cubebene 204.35 g/mol Aerial parts
Maceration 24 h, GC-MS
Hydrodistillation, solvent
extraction, GC-MS
[35]
[36]
45. β-Caryophyllene 204.35 g/mol Aerial parts
Maceration 24 h, GC-MS
Hydrodistillation, GC-MS
SFME, hydrodistillation 1 h, GC-MS
[35]
[38]
[37]
Oxygenated Sesquiterpene
46. α-Bisabolol 222.37 g/mol Aerial parts Hydrodistillation 4 h, GC-MS [28]
47. α-Cadinol 222.37 g/mol Aerial parts Hydrodistillation 4 h, GC-MS [28]
48. Nerolidol 222.37 g/mol Leaf
SFME, hydrodistillation 1 h, GC-MS
[37]
49. Caryophyllene oxide 220.35 g/mol Aerial parts Maceration 24 h, GC-MS [35]
Other Compounds
50. trans-4-
Methoxycinnamaldehyde 162.18 g/mol Leaf Hydrodistillation, GC-MS [34]
51.
Mandelic Acid (Benzeneacetic
acid, alpha.-hydroxy) 152.15 g/mol Leaf Hydrodistillation, GC-MS [34]
52. Phenylethanolamine 137.18 g/mol Leaf Hydrodistillation, GC-MS [34]
53. N-Benzyl-N-ethyl-p-
isopropylbenzamide 281.4 g/mol Leaf Hydrodistillation, GC-MS [34]
54. cis-2-(2-pentenyl) furan 136.19 g/mol Leaf Maceration, GC-MS [32]
SFME: Solvent-free microwave extraction.
Plants 2023, 12, x FOR PEER REVIEW 6 of 26
51. Mandelic Acid (Benzeneacetic acid,
alpha.-hydroxy) 152.15 g/mol Leaf Hydrodistillation, GC-MS [34]
52. Phenylethanolamine 137.18 g/mol Leaf Hydrodistillation, GC-MS [34]
53. N-Benzyl-N-ethyl-p-isopropylben-
zamide 281.4 g/mol Leaf Hydrodistillation, GC-MS [34]
54. cis-2-(2-pentenyl) furan 136.19 g/mol Leaf Maceration, GC-MS [32]
SFME: Solvent-free microwave extraction.
Figure 2. Structure of monoterpenoids.
Figure 2. Structure of monoterpenoids.

Plants 2023,12, 4148 7 of 25
Plants 2023, 12, x FOR PEER REVIEW 7 of 26
OH
OH
HO
OO
O
OH
O
OH
N
H
HO
O
NO
26 27 28 29 30
31 32 33 34 35 36
37 38 39 40 41 42
43 44 45 46 47
48 49 50 51
52 53 54
Figure 3. Structure of sesquiterpenoids.
5. Ethnomedicinal Evidence for O. basilicum L.
More than 10% of plant species and over 50,000 species have been utilized for the
development of medications and healthcare products [2]. O. basilicum L., also known as
the king of herbs in the Greek word, has been recognized since ancient times for its ther-
apeutic properties and was used in the Unani and Ayurvedic medical systems [41]. The
proper way to use medicinal plants is typically passed down from one generation to the
next and often pertains to traditional remedies for age-old ailments [42]. These traditional
beliefs about medicinal plants will be enhanced by the integration of technology for the
production of sustainable pharmaceuticals.
Ethnobotany is a subdiscipline of ethnobiology, which studies the traditional botan-
ical knowledge in different cultures, the techniques for utilizing plants, the management
of plant resources, and the role, which plants play in ritual, cultural, or religious beliefs
[43]. As a result, it serves as a foundation for selecting plants, which can be developed for
medicinal purposes [44]. Ethnobotany contributes to exploration of the ways to fill the gap
between scientific research and cultural or indigenous understanding [45]. The majority
of ethnobotanical and ethnopharmacological studies have been conducted to acquire
knowledge about the use of medicinal plants to treat various illnesses [46].
Figure 3. Structure of sesquiterpenoids.
5. Ethnomedicinal Evidence for O. basilicum L.
More than 10% of plant species and over 50,000 species have been utilized for the
development of medications and healthcare products [
2
]. O. basilicum L., also known as the
king of herbs in the Greek word, has been recognized since ancient times for its therapeutic
properties and was used in the Unani and Ayurvedic medical systems [
41
]. The proper way
to use medicinal plants is typically passed down from one generation to the next and often
pertains to traditional remedies for age-old ailments [
42
]. These traditional beliefs about
medicinal plants will be enhanced by the integration of technology for the production of
sustainable pharmaceuticals.
Ethnobotany is a subdiscipline of ethnobiology, which studies the traditional botanical
knowledge in different cultures, the techniques for utilizing plants, the management of
plant resources, and the role, which plants play in ritual, cultural, or religious beliefs [
43
].
As a result, it serves as a foundation for selecting plants, which can be developed for
medicinal purposes [
44
]. Ethnobotany contributes to exploration of the ways to fill the gap
between scientific research and cultural or indigenous understanding [
45
]. The majority

Plants 2023,12, 4148 8 of 25
of ethnobotanical and ethnopharmacological studies have been conducted to acquire
knowledge about the use of medicinal plants to treat various illnesses [46].
India is known as a country for the knowledge and utilization of herbal medicine [
47
].
Indigenous and local people in the Bageshwar District of Uttarakhand, India, use O.
basilicum L. leaf and seed for treating fever, cough, and cold [
48
]. The community around
Lawachara National Park in Bangladesh utilizes O. basilicum L. leaf as a treatment for
reducing high blood pressure, fever, and cough [
49
]. Another area in Lalmohan, Bhola
District, Bangladesh, utilized the whole plant of O. basilicum L. to treat fever and as a
carminative [50].
Remote areas in India, such as the Uttara Kannada District, used herbal treatments
with O. basilicum L. to treat reproductive diseases, such as dysmenorrhea, by crushing the
bark in milk and drinking it once a day for seven days [
51
]. Additionally, local healers from
the Khatling Valley and Pauri District in Uttarakhand, India, drink the decoction of leaves
and seeds of O. basilicum L. for fever, cough, cold, and urinary problems [
52
,
53
]. Traditional
healers in the North West Ganjam District, Odisha, India, employ powder and decoction
from the leaves of O. basilicum L. for treating dysuria, cough, and cold [
54
]. Meanwhile, the
traditional healers from the Rabha Tribe in the Kamrup District, India, use the leaves and
inflorescence as a remedy for cough and chronic fungal infections [55].
Pakyoung in East Sikkim, India, reported that the leaves and seeds of O. basilicum
L. were utilized for colds, coughs, fevers, and constipation [
56
]. Based on an interview
with 33 traditional healers from Nelliyampathy, Kerala, India, the leaves and seeds of
O. basilicum L. were made into paste, inhalation, juice, and infusion, which could treat
tumors, headaches, insomnia, heart trembling, coughs, chest pain, dysentery, diarrhea, and
gonorrhea [57].
In Dharan, Nepal, O. basilicum L. leaf juices are used for digestive disorders, such
as diarrhea, dysentery, constipation, gastritis, and vomiting [
58
]. In the Sulaymaniyah
province in Iraq, it had been reported that the local community applied O. basilicum L. as an
ethnobotanical treatment for headaches, colds, and halitosis (bad breath) [
59
]. Meanwhile,
in the Peshawar Valley, Pakistan, O. basilicum L. leaves were made into an extract and given
orally to help improve digestion and for other purposes, such as ornamental decoration [
60
].
O. basilicum L. has a history of being used as a medical treatment in some southeast
Asian nations for a variety of diseases. Traditional healers from Phatthalung, southern
Thailand, use O. basilicum L. for treating flatulence and peptic ulcers [
61
]. Local tribes from
Tina and Libas Gua Village, Mindanao, Philippines, utilized the leaves by rubbing them
around the affected area of the body to treat cold sores [
62
]. Furthermore, in Indonesia,
this plant is widely used in the big islands, such as Sulawesi, Sumatra, and Kalimantan.
The tribes in Kolaka and East Kolaka, Southeast Sulawesi, make use of the leaves of O.
basilicum L. for treating tuberculosis [
63
]. Meanwhile, the traditional healers in many
areas of Sumatra utilized the seeds of O. basilicum L. as a therapy for treating back pain
sickness [
64
]. The sub-ethnic Dayak tribe—like the Dayak Linoh tribe, who live in the
Sintang District, West Kalimantan—utilized the leaves, flowers, and fruit to reduce body
odor and fever [
65
]. Another sub-ethnic group, Dayak Tamambaloh, in the Kapuas Hulu
District, West Kalimantan, used the leaves to treat ringworm, blisters, and reduce body
odor [66].
In addition, in the South American continent, e.g., in Maragogipe, State of Bahia,
Brazil, the leaves were made into tea for the treatment of delayed menstruation, fever, flu
in children, indigestion, and nasal congestion [
67
]. In an ethnomedicine inventory record
in the African continent, e.g., the Abia State in Nigeria, the seeds of O. basilicum L. were
used for treating diarrhea [
68
]. In Western Oromia State, Ethiopia, this species is used to
cure allergic reactions by crushing it and mixing it with food [
69
]. In central Kenya, the
plant is used for curing common cold [70]. In the Rainforest Research Station, Ondo State,
Nigeria, the whole plant of O. basilicum L. is used for treating inflammation [71].
The utilization of O. basilicum L. as a herbal remedy has been observed on a global
scale and spread across regions such as Asia, Africa, and South America. Traditional healers

Plants 2023,12, 4148 9 of 25
from various regions used the whole plant, as well as some parts of the plant, such as leaves,
seeds, inflorescences, fruits, and stems. The plants were made into infusion, inhalation,
paste formulations, powder, tea, and they were incorporated into food. According to the
beliefs of traditional healers, O. basilicum L. possesses therapeutic properties in the man-
agement of typical illnesses affecting the digestive, respiratory, urinary, and reproductive
systems. Additionally, certain individuals within the society employed this particular plant
for the purpose of enhancing decorative esthetics.
6. Antimicrobial Activities and Biomedical Uses
Medicinal plants contain rich, yet underexploited, bioactive compounds, with a limited
amount of their potential qualities having been thoroughly examined [
72
]. The exploration
of a medicinal plant with high chemical constituents holds promise in the pursuit of de-
veloping therapeutic treatments in the future [
73
]. Each part of the plant offers potentially
valuable biomedical knowledge, which remains uncovered [
74
]. As one of the medicinal
plants, O. basilicum L. possesses considerable undiscovered potential in the field of antimi-
crobial and biomedical research. The goal of biomedical therapy through the utilization of
medicinal plants requires continuous exploration and development.
6.1. Antiviral Activity
Based on the ethnomedicinal records and data, traditional healers all over the world
consider O. basilicum L. to be an important herb. A number of studies have shown that
O. basilicum L. has the potential for antiviral activity. For instance, a recent study of O.
basilicum L. against SARS-CoV-2 with an in silico assay showed that polyphenol con-
stituents apigenin-7-glucuronide and dihydrokaempferol-3-glucoside have binding affinity
for
−
8.77 Kcal/mol and
−
8.96 Kcal/mol, respectively, which possess great potential for
antiviral activity. These compounds have binding affinity with the main protease (M
pro
)
enzymes on SARS-CoV-2 [
75
]. The M
pro
enzyme, typically referred to as 3-chymotrypsin-
like protease (3CL
pro
), serves a pivotal role in viral replication and is being targeted as a
way of preventing COVID-19 infection [76].
Aside from the polyphenol compound, for the first time, monoterpenes showed the
potential for antiviral activity toward SARS-CoV-2. An
in vitro
study demonstrated that five
different monoterpene compounds, such as carvone, carvacrol, menthofuran, 1,8 cineole,
and pulegone, potentially inhibited SARS-CoV-2 in infected Vero 76 cells. Among these five
compounds, carvacrol and carvone showed significant antiviral activity with half-maximal
inhibitory concentration (IC
50
) < 100
µ
M. In addition, an essential oil, which contained the
highest carvone concentration (>200 mg/mL), had the greatest antiviral activity with IC
50
127
±
4.63 ppm. The antiviral properties of monoterpene compounds had been observed
to bind and interrupt the important viral proteins, among them, the main protease and
spike protein [
77
]. On top of its potent antiviral effects, carvacrol had a favorable safety
profile, suggesting its potential as a viable candidate for the development of preventive
therapies [78].
Another recent study tested the lipophilic fraction of the stem of this plant
in vitro
and in silico against dengue virus (DENV). This
in vitro
study revealed that the fraction
significantly reduced DENV titer in pre-treatment and post-treatment conditions at a
concentration of 3.125
µ
g/mL. Meanwhile, an in silico study of the lipophilic fraction
showed that the two compounds had great binding affinity. Stigmasterol had a binding
affinity for
−
8.3 Kcal/mol with NS1 protein, and campesterol exhibited the biggest binding
affinity for
−
8.2 Kcal/mol with E glycoprotein [
33
]. A computational study demonstrated
that this plant has the potential for antiviral drug development through its mechanism
of inhibiting the active site of human immunodeficiency virus (HIV) gp120 and gp41.
α
-guaiene is the compound with the highest negative value for binding affinity, which
was
−
9.62 Kcal/mol at the active site gp120, and sitosterol displayed a binding affinity for
−
10.99 Kcal/mol at the active site gp41 [
79
]. Furthermore, an
in vitro
study of the crude

Plants 2023,12, 4148 10 of 25
ethanolic extract of O. basilicum L. leaves demonstrated antiviral activity against Zika virus
(ZIKV) with 97% virus infectivity at the highest concentration (1:16 dilution) [80].
There are a few studies, which show that O. basilicum L. can treat some viruses infecting
livestock, such as cattle and poultry. An
in vitro
study of alcoholic extract from O. basilicum
L. leaves showed potential in managing the Newcastle disease virus (NDV), which infects
poultry, with a reduction titer up to 10
−7
at the concentration of 500
µ
g/mL [
81
]. Kubiça
et al. reported an
in vitro
study of 1,8-cineole and camphor, which both reduced the
plaque in bovine viral diarrhea virus (BVDV) by approximately 75% and 84%, respectively,
at the maximum non-toxic dose [
82
]. Another ethanolic extract of O. basilicum L. was
made into an ointment for daily application in bovine cutaneous papillomatosis, which is
caused by bovine papillomavirus (BV). The ointment was made into a 2% formulation with
20 mg/g weight per weight (w/w) of ethanolic extract and white petroleum jelly for the
base. Clinically, the papilloma began to regress and eventually disappeared between days 7
and 21, and the skin texture progressively returned to normal. The antiviral activity of this
topical formulation may be due to the phenolic, flavonoid, tannin, and alkaloid compounds
found in it [83].
Another crude aqueous and ethanolic extracts of the whole plant of O. basilicum L., as
well as selected purified constituents, exhibited antiviral activity against DNA viruses, such
as the herpes virus (HSV), adenoviruses (ADV), hepatitis-B virus (HBV), and RNA viruses,
such as coxsackievirus B1 (CVB1) and enterovirus 71 (EV71) [
84
]. This
in vitro
research
found that purified constituents, such as apigenin, ursolic acid, and linalool, had antiviral
activity against HSV-1 similar to that of acyclovir and also against HBV and enterovirus.
The strongest purified constituents were ursolic acid against HSV-1 (EC
50
6.6 mg/L),
ADV-8 (EC
50
4.2 mg/L), CVB1 (EC
50
0.4 mg/L), and EV71 (EC
50
0.5 mg/L). Furthermore,
apigenin possessed the highest antiviral activity against HSV-2 (EC
50
9.7 mg/L), ADV-3
(EC
50
11.1 mg/L), and hepatitis-B surface antigen (EC
50
7.1 mg/L). Meanwhile, linalool has
recorded moderate anti-adenoviral activity against ADV-3, ADV-8, and it has exhibited the
strongest effects against ADV-11 (EC
50
16.9 mg/L) [
84
]. In addition, methanolic extracts of
O. basilicum L. significantly inhibited herpes simplex virus 1 strain F (HSV-1F) after viral
adsorption [
85
]. Another purified compound, such as eugenol, from the methanolic extract
of O. basilicum L. showed inhibition of pre-HIV-1 infection in the host cell at the effective
concentration of 350 µg/mL [86].
The evidence presented above indicates that the phytochemical constituents of O.
basilicum L. hold the potential to be developed into new drugs, offering an opportunity to
address issues such as drug resistance and side effects [
87
]. Understanding the phytochem-
ical constituent mechanisms of action in pharmacology is crucial before developing drugs
from O. basilicum L. [
88
]. The antiviral properties of phytochemicals have been subject of
substantial research in recent years, even after the world experienced a worldwide pan-
demic [
89
]. Therefore, it is crucial to maintain and expand this research into their antiviral
properties in light of ongoing global health challenges.
6.2. Antibacterial Activity
The increasing incidence of antibiotic resistance in recent years has prompted an
immediate demand for new strategies and innovative antibiotic formulations [
90
]. The
aforementioned issue arises from the irresponsible use of antibiotics in the context of
human healthcare and the practice of animal husbandry [
91
]. O. basilicum L. is one of many
medicinal plants, which have demonstrated potential as antibacterial agents. Essential oils,
methanolic extracts, and fractions from the plant have been explored over these past few
years for their antibacterial properties against Gram-positive and Gram-negative bacteria.
The evidence of antibacterial activity against Gram-positive and Gram-negative bacteria is
shown at the minimum inhibitory concentration (MIC) in Table 2, and the diameter of the
inhibitory zone is shown in Table 3.

Plants 2023,12, 4148 11 of 25
Table 2. Minimum inhibitory concentration (MIC) value of antibacterial activity of O. basilicum L.
Bacterial Species Essential Oil/Extract MIC Value Reference
Gram Positive
Bacillus cereus (ATCC 11778) Essential oil
Essential oil and methanolic extract
10.80 µL/mL
62.5 µg/mL
[92]
[93]
Bacillus subtilis Essential oil and methanolic extract 125 µg/mL [93]
Bacillus megaterium Methanolic extract 62.5 µg/mL [93]
Enterococcus faecalis (ATCC 19433) Essential oil 0.75 mg/mL [94]
Listeria monocytogenes Essential oil and methanolic extract 125 µg/mL [93]
Micrococcus luteus (ATCC 10240) Essential oil 0.50 mg/mL [94]
Sarcina sp. Essential oil 0.75 mg/mL [94]
Staphylococcus aureus (ATCC 6538P)
Essential oil
Essential oil
Essential oil
Essential oil and methanolic extract
2.45 µL/mL
32 µg/mL
1 mg/mL
62.5 µg/mL
[92]
[95]
[94]
[93]
Staphylococcus epidermidis Essential oil 0.75 mg/mL [94]
Streptococcus mutans Essential oil 0.75 mg/mL [94]
Gram Negative
Acinetobacter sp. Essential oil 0.75 mg/mL [94]
Aeromonas sp. Essential oil 1 mg/mL [94]
Citrobacter freundii (ATCC 8090) Essential oil 1 mg/mL [94]
Escherichia coli (ATCC 25922) Essential oil
Methanolic extract
10.80 µL/mL
125 µg/mL
[92]
[93]
Klebsiella pneumoniae (ATCC 13833) Essential oil 0.75 mg/mL [94]
Proteus mirabilis (ATCC 25933) Essential oil 1 mg/mL [94]
P. vulgaris (ATCC 13315) Essential oil 0.75 mg/mL [94]
Pseudomonas aeruginosa (ATCC 27853)
P. aeruginosa (ATCC 25853)
P. aeruginosa (1662339)
Essential oil
Essential oil
Essential oil
22.68 µL/mL
256 µg/mL
32 µg/mL
[92]
[95]
[95]
Salmonella choleraesuis (ATCC 10708) Essential oil 0.5 mg/mL [94]
Salmonella typhimurium (ATCC 14028) Essential oil 22.68 µL/mL [92]
Serratia marcescens (ATCC 13880) Essential oil 0.25 mg/mL [94]
Shigella boydii Essential oil 250 µg/mL [93]
Shigella dysenteriae Essential oil and methanolic extract 250 µg/mL [93]
Shigella flexneri (ATCC 12022) Essential oil 0.75 mg/mL [94]
Vibrio parahaemolyticus Essential oil 250 µg/mL [93]
Vibrio mimicus Essential oil 250 µg/mL [93]
Yersinia enterocolitica (ATCC 10460) Essential oil 0.25 mg/mL [94]
Table 3. Diameter of zone inhibition of antibacterial activity of O. basilicum L.
Bacterial Species Essential Oil/Extract Diameter of Zone Inhibition Reference
Gram Positive
Bacillus cereus Essential oil
Ethyl acetate fraction
25 mm
21.1 mm
[96]
[93]
Bacillus subtilis Ethyl acetate fraction
Methanolic extract
19.3 mm
31.86 mm
[93]
[97]
Bacillus megaterium Ethyl acetate fraction 18.2 mm [93]
Clostridium perfringens type C Methanolic extract 31.13 mm [97]
Cutibacterium acnes (ATCC 11827) Essential oil 18.13 mm [38]
Enterococcus sp. Methanolic extract 30.73 mm [97]
Enterococcus faecalis (ATCC 19433) Essential oil
Essential oil
10.3 mm
11.2 mm
[94]
[98]
Listeria monocytogenes Essential oil 17.1 mm [93]

Plants 2023,12, 4148 12 of 25
Table 3. Cont.
Bacterial Species Essential Oil/Extract Diameter of Zone Inhibition Reference
Micrococcus luteus (ATCC 10240) Essential oil 13.5 mm [94]
Sarcina sp. Essential oil 14.6 mm [94]
Staphylococcus aureus (ATCC 6538)
S. aureus (ATCC 6538)
S. aureus (ATCC 25923)
S. aureus
Essential oil
Ethyl acetate fraction
Essential oil
Methanolic extract
9 mm
17.1 mm
9.7 mm
30.66 mm
[96]
[93]
[98]
[97]
Staphylococcus epidermidis (ATCC 12228)
Essential oil 13.3 mm [98]
Staphylococcus mutans (ATCC 25175) Essential oil 11 mm [94]
Gram Negative
Acinetobacter sp. Essential oil 15 mm [94]
Aeromonas sp. Essential oil 10.6 mm [94]
Citrobacter freundii (ATCC 8090) Essential oil 11.6 mm [94]
Escherichia coli
E. coli
E. coli
E. coli
E. coli (ATCC 25922)
Essential oil
Essential oil
Ethyl acetate fraction
Methanolic extract
Essential oil
11 mm
10.3 mm
14.2 mm
28.30 mm
13.5 mm
[96]
[94]
[93]
[97]
[98]
Klebsiella pneumoniae
Essential oil
Essential oil
Methanolic extract
12.2 mm
17.2 mm
26.66 mm
[94]
[98]
[97]
Proteus mirabilis (ATCC 25933) Essential oil
Essential oil
11.3 mm
13.1 mm
[94]
[98]
Proteus vulgaris (ATCC 13315) Essential oil 18 mm [94]
Pseudomonas aeruginosa Methanolic extract 28.83 mm [97]
Salmonella choleraesuis (ATCC 10708) Essential oil 10 mm [94]
Salmonella typhymurium Essential oil
Methanolic extract
10 mm
15.30 mm
[96]
[97]
Serratia marcescens (ATCC 13880) Essential oil
Essential oil
16.6 mm
10.4 mm
[94]
[98]
Shigella boydii Essential oil 13.3 mm [93]
Shigella dysenteriae Ethyl acetate fraction 15.2 mm [93]
Shigella flexneri (ATCC 12022) Essential oil 17.1 mm [94]
Vibrio parahaemolyticus Ethyl acetate fraction 16.2 mm [93]
Vibrio mimicus Methanolic extract 51.2 mm [93]
Xanthomonas sp. Methanolic extract 14.36 mm [97]
Yersinia enterocolitica (ATCC 10460) Essential oil 12.6 mm [94]
An
in vitro
study of the antibacterial characteristics of O. basilicum L. essential oil
demonstrated inhibition and eradication activities against Vibrio strains’ mature biofilm.
A concentration of 50 mg/mL of O. basilicum L. essential oil was shown to greatly inhibit
the biofilm, with a percentage of 55% for V. parahaemolyticus and up to 87.45% for both
V. vulnificus and V. cholerae. The bactericidal effects of this essential oil may have a cor-
relation with the high amount of linalool found in its composition [
99
]. The potential
antibacterial activity of monoterpene compounds involves the degradation of cell walls
and cell membranes, as well as the interruption of membrane protein and ion transport
processes [100].
Another study demonstrated that methanolic leaf extracts of O. basilicum L. were shown to
have a great potential in antibacterial activity against B. cereus,P. aeruginosa,L. monocytogenes,E.
coli,M. flavus, and S. aureus, with MIC < 0.5 mg/mL and
MBC < 0.9 mg/mL
. This methanolic
extract contains some polyphenol compounds, such as 3,4-dihydroxyphenylacetic acid and
rutoside (rutin), which mainly contribute to the antibacterial effects [
30
]. The antibacterial
effects of polyphenol compounds arise from their ability to interact with bacterial cell
walls and membranes, disrupt protein regulation, inhibit microbial enzymes, and exhibit
iron-chelating capabilities [101].

Plants 2023,12, 4148 13 of 25
The essential oil of O. basilicum L. combined with the antibiotic imipenem had a
synergistic interaction, resulting in antibacterial activity against S. aureus and P. aeruginosa.
Meanwhile, the essential oil combined with the antibiotic ciprofloxacin had an antagonistic
and indifferent interaction [
95
]. This means that O. basilicum L. phytochemical-derived
substances have the potential to enhance the existing antibiotics. Anwar et al. demonstrated
that O. basilicum L. in multiple regions of Saudi Arabia exhibited variations in antibacterial
activity because of the different chemical compositions, which are influenced by the diverse
agro-climatic regions [102].
In addition to diverse regions, various factors influence antimicrobial activity, such
as the chemical compounds, bacterial strain, temperature, and bacterial cell number [103].
The evidence of antibacterial properties indicates that O. basilicum L. has a broad spectrum
of antibacterial activity against both Gram-positive and Gram-negative bacteria.
6.3. Antifungal Activity
In the intensive care unit (ICU), invasive fungal infections are commonly identified
in patients with a weakened immune system [
104
]. Fungal infections are a major cause of
infectious-disease-related mortality around the world [
105
]. Additionally, because fungi
are eucaryote organisms, they only have a few molecular targets, which can be exploited
by medications to activate their effects [
106
]. Recently, phytochemical constituents have
received a lot of interest in the research and development of antifungal medications.
The essential oil of O. basilicum L. exhibits antifungal activity against pathogenic
fungal Aspergillus flavus at a concentration of 1000 ppm, which could suppress the fungal
growth and aflatoxin B1 biosynthesis. The chemical compounds, which possess good
antifungal properties, are linalool and 1,8-cineol from monoterpene and eugenol from
polyphenol [
40
]. The commercial basil extracts in Slovenia were tested against several
Fusarium species for antifungal properties. The extracts were found to inhibit colony growth
in F. proliferatum at concentrations of, respectively, 0.35% and 0.70% by up to 33.37% and
44.30%, and they inhibited colony growth in F. subglutinans by up to 24.74% and 29.27%.
The commercial extracts are known to contain estragole (86.72%), trans-
α
-bergamotene
(2.91%), and eucalyptol or 1,8-cineole (2.67%) [107].
The essential oil of O. basilicum L. has antifungal properties against Candida albicans,
with an inhibition zone of 27 mm. These activities are supported by linalool, methyl
chavicol,
β
-elemene, and
α
-bulnesene [
108
]. In Serbia, twelve cultivars of O. basilicum L.
were tested against seven species of fungi, including A. ochraceus,A. versicolor,A. fumiga-
tus,A. niger,Penicillium funiculosum,P. ochrochloron, and Trichoderma viridae. The twelve
cultivars exhibited great antifungal properties in inhibiting fungal growth, with the mini-
mum inhibition concentration being 10–100-fold higher than the commercial antifungal
drug ketoconazole and minimum fungicidal concentrations ranging from 0.14
µ
g/mL to
27.00 µg/mL
. These antifungal properties of the twelve cultivars of O. basilicum L. may be
due to high linalool composition in the essential oils [109].
In addition to essential oil, the methanolic fraction from the aerial parts of O.basilicum
L. showed antifungal activity. The fraction was tested against eight species of fungi,
including A. flavus,A. niger,Penicillium,Rhizopus solani,Alterneria alternata,Candida albicans,
Curvilaria lunata, and A. fumigates. These methanolic fractions exhibited strong inhibition of
fungal growth, from the lowest at 10% up to 100%, at concentrations of 1 mg/mL, 3 mg/mL,
and 6 mg/mL. C. albicans was one of the resistant species; at 3 mg/mL, it only had 17%
inhibition compared to other species, which had 43–100% inhibition [
110
]. In contrast, the
ethanolic extracts from the aerial parts of O. basilicum L. demonstrated antifungal activities
against C. albicans. The extracts showed the presence of an inhibitory zone measuring
18 mm for the fungi [111].
A study showed that high concentrations of terpene compounds, such as citral,
eugenol, nerolidol, and
α
-pinene, demonstrated an antifungal mechanism by breaking
down the cell membrane [
112
]. Meanwhile, an in silico study of polyphenols from the
plant, including rutin, kaempferol, and quercetin, demonstrated an underlying antifungal

Plants 2023,12, 4148 14 of 25
mechanism. The polyphenol compound was found to have the ability to inhibit fungal
enzymes. Rutin—also called rutoside—had the greatest antifungal activity through binding
with 14-alpha demethylase (CYP51) and nucleoside diphosphokinase (NDK), with binding
affinity for −9.4 and −8.9, respectively [113].
The synergistic action of various constituents comprising the essential oil suppresses
the chance of resistance. This is attributed to the difficulty the pathogens face in adapting
resistance characteristics against multiple compounds present in essential oils [
114
]. This
indicates that O. basilicum L. has potential for development into a new class of natural
antifungal drugs.
6.4. Biomedical Activity
Numerous records and findings have demonstrated the advantages of O. basilicum
L. plant in the medical field, such as its antioxidant, anticancer, analgesic, antidiabetic,
anti-inflammatory, and antidepressant effects [
12
]. The O. basilicum L. essential oil from the
seeds had good antioxidant activity when using the DPPH assay compared to the positive
control, which was the Trolox compound. Great antioxidant activity was shown with an
inhibition concentration (IC
50
) of 23.44
±
0.9
µ
g/mL [
115
]. Antioxidant activity eventually
resulted from the synergy of each compound and had correlation with the total phenolic
compound, which composed the essential oil [
116
]. The phenolic compound has the ability
to donate a hydrogen atom to the free radicals, the ability to chelate metal cations, and the
ability to scavenge free radicals [
39
,
117
]. Stanojevic et al. also reported that with the DPPH
assay, basil essential oil had good antioxidant properties with effective concentration (EC
50
)
at 2.38
±
0.10 mg/mL, and it could be used as an alternative to synthetic antioxidants
with a higher safety profile [
108
]. In addition to essential oil, the O. basilicum L. hexane
extracts also showed possible antioxidant activity in a concentration- and dose-dependent
manner [118].
The O. basilicum L. in Jordan was tested against three cancer cell lines for anti-
cancer activity, such as the triple-negative breast cancer cell line (MDA-MB-231) with IC
50
432.3 ±32.2 µg/mL
, the ER+ breast cancer cell line (MCF7) with IC
50 320.4 ±23.2 µg/mL
,
and the glioblastoma cancer cell line (U-87 MG) with IC
50
431.2
±
15.3
µ
g/mL. It turned
out that the essential oils containing major components of linalool, eugenol, and eucalyptol
exhibited potential anticancer activity [
119
]. The essential oil was also tested on cancer
cell lines from liver cancer (Hep 3B) and breast cancer (MCF-7), which resulted in good
cytotoxic effect on both cell lines [
115
]. In addition to the essential oils, methanolic ex-
tracts from the aerial parts of the plant also exhibited promising anticancer activity against
MCF-7 and MDA-MB-231. The anticancer properties are shown by the expression level
of apoptosis-related genes, which decreases the bcl-2 gene’s ability to act as an inhibitor
protein for programmed cell death and allows the cell to undergo apoptosis. In this study,
the anticancer activity of O. basilicum L. came from the active compound eugenol [120].
The combination of Morus nigra and O. basilicum L. extracts was tested in various cancer
cell lines and normal human cells for anticancer activity. It was discovered that the chloro-
form extracts (MO2C) possessed the highest anticancer activity. The reason for this is that
MO2C was cytotoxic against all tested cell lines at the lowest
concentration—particularly
the breast cancer cell line—and had selective cytotoxicity toward the normal cell line. In
addition, this extract contains
α
-trans-bergamotene, germacrene D, selin-4,7(11)-diene,
2-decel-1-ol, and 2-tridecen-1-ol, which play anticancer roles. The anticancer capacities
observed in cell morphology include shrinkage, loss of cellular integrity, cell detachment,
and contraction of the cytoplasm [121].
Another
in vitro
study demonstrated that the methanolic, hexane, and dichloromethane
extracts showed a potential antidiabetic property. These three extracts were subjected to a
cytotoxic assay, which is considered safe for methanolic extracts up to 0.25 mg/mL and
for hexane and dichloromethane extracts up to 0.5 mg/mL. The hexane extract demon-
strated an “insulin-like” effect in the absence of insulin due to translocation of the glu-
cose transporter (GLUT4) to the plasma membrane. This study stated that there were

Plants 2023,12, 4148 15 of 25
17 newly identified compounds, which possibly played antidiabetic roles in the extracts.
Some of these compounds contained glycerol, cyanuric acid, talose, oleamide, inositol,
hydroquinone-beta-d-glucopyranoside, pentane-1,2,5-triol, and glucopyranose. Inositol
was first found in the O. basilicum L. methanolic extract in this study [122].
Meanwhile, an
in vivo
study performed in male albino mice showed that O. basilicum
L. leaf extract administered orally had the potential to improve neuromuscular coordina-
tion, active behavior, the ability to recognize objects, and short-term memory. The optimum
daily supplementation dose was found to be 100 mg/mL solvent/kg body weight and
was considered suitable for oral administration without any safety concerns [
123
]. Hy-
droethanolic extract, ethyl acetate, and n-hexane fractions had anticonvulsant and neu-
roprotective characteristics, which prevented oxidative damage to the brain tissue, with
optimum dose at 200 mg/kg [
124
]. For the first time, new compounds called 5,7-dihydroxy-
3
0
,4
0
,5
0
-trimethoxyflavone and 3-hydroxy-3
0
,4
0
,5
0
-trimethoxyflavone have been found in
O. basilicum L. leaf extracts and fractions. An in silico study showed that both compounds
had binding interaction energy for
−
9.93309 and
−
15.9683, respectively, with Caspase-3
target protein. Both of these compounds helped improve long-term memory by reducing
Caspase-3 concentration and suggesting the role of anti-apoptotic cells against neuron cells.
This neuroprotective ability is due to the combination of anticholinergic, antioxidant, anti-
inflammatory, and anti-apoptotic effects of the compound [
125
]. The inhalation of essential
oil derived from O. basilicum L. has been shown to possess neuroprotective properties and
exhibit depressive effects in mice. The essential oil demonstrated antidepressant properties
in mice subjected to chronic unpredictable mild stress [126].
Moreover, O. basilicum L. has been identified as an effective agent in exerting anti-
inflammatory effects. An
in vivo
study performed in mice showed that the essential oil
of O. basilicum L. with the estragole (methyl chavicol) chemotype in doses of 100 mg/kg
and 50 mg/kg greatly reduced paw edema induced by carrageenan by 74% and 44%,
respectively, between the first and fifth hour of evaluation. Furthermore, these doses of
essential oil are deemed safe for oral administration [
127
]. An
in vitro
study showed that
O. basilicum L. treated with chemical elicitors, such as arachidonic acid, jasmonic acid, and
β
-aminobutyric acid, enhanced the flavonoid and phenolic content, which possess anti-
inflammatory properties. This finding showed that a plant with arachidonic acid elicitor
had the greatest inhibitory effect against lipoxygenase (LOX) (EC
50
=
1.67 mg FW mL−1
)
and cyclooxygenase (COX) (EC
50
= 0.31 mg FW mL
−1
). The inhibitory efficacy exhibited
positive correlation with the increased content of rosmarinic, benzoic, and o-coumaric
acids [
128
]. Moreover, an
in vitro
study demonstrated that ethanolic leaf and leaf callus
extracts significantly reduced nitric oxide as pro-inflammatory mediators with concentra-
tions of 0.01–1 mg/mL on RAW 264.7 macrophage cells. This anti-inflammatory activity
may be a result of the major compounds found in the extracts, which are 2,3-dihydroxy-3,5-
dihydroxy-6-methyl-4H-pyran-4-one and 2-methoxy-4-vinylphenol [129].
Another
in vivo
study demonstrated that O. basilicum L. methanolic extract emulgel
formulation represented a potential alternative for second-degree-burn wound-induced
rabbits. Formulations with 5% extract, polymer, and other excipients were compatible
and had a good safety profile for topical emulgel. This extract formulation showed a
healing capacity on the 16th day, with 98.78% wound contraction, which was insignificantly
different to the healing capacity of commercial healing products [130].
In addition, a novel hydrogel formulation based on O. basilicum L. and Trifolium
pratense extract combination had a great wound healing ability.
In vitro
tests demonstrated
that the combined extract with concentration of 50
µ
g/mL had the greatest healing efficacy
in terms of complete healing time and fibroblast density. An
in vivo
study also showed
that the combined extract healed the wound 100% on the 13th day—better than the control
group—which means that it exhibited a remarkable wound healing capability. Moreover,
the hydrogel formulation was tested in a clinical case of a patient with Psoriasis vulgaris
twice a day. The formulation was shown to reduce erythema symptoms within one week of
treatment. The tremendous wound healing capability could be explained by the synergistic

Plants 2023,12, 4148 16 of 25
effect of the extract’s phytochemical mixture. The O. basilicum L. extract was rich in
phenolic and flavonoid contents, especially ferulic acid; meanwhile, the T. pratense was rich
in chlorogenic acid [
131
]. Ferulic acid is known for its ability to enhance wound healing
through the promotion of angiogenesis, reduction in oxidative stress, and inhibition of
bacterial growth [132].
Moreover, there are three novel compounds found in O. basilicum L., such as inositol,
5,7-dihydroxy-3
0
,4
0
,5
0
-trimethoxyflavone, and 3-hydroxy-3
0
,4
0
,5
0
-trimethoxyflavone. The
studies above indicated that O. basilicum L. holds significant potential for development
and formulation into a natural pharmaceutical alternative. Thus, additional investigation
of the drug delivery system and clinical trials are necessary in order to create natural
accessible medication.
7. Biotechnological Development in O. basilicum L. Research
The field of biotechnology involves the utilization of scientific methodologies to alter
and enhance the characteristics of plants, animals, and micro-organisms in order to increase
their overall value [
133
]. The demand for herbal medicine on a global scale is substantial
and exhibits a consistent growth rate. Various technologies have been implemented to
facilitate the promotion of bioactive compounds in medicinal plants [
134
]. Secondary
metabolites, which are considered vital constituents of the plants, hold significant economic
value due to their applications as pharmaceutical products, perfumes, pigments, and food
additive products [135].
7.1. Green Nanotechnology Production in O. basilicum L. for Medical Application
Previously, we discussed the antibacterial and antifungal properties of O. basilicum L.,
which were extensively explored from 2010 to 2018, revealing its potential for combating
various bacterial and fungal infections. Over the last five years, numerous studies on
nanotechnology have demonstrated the ways of enhancing the antimicrobial properties of
this plant. One notable advantage of using plant extracts for synthesizing nanoparticles
is their ability to generate a larger zone of inhibition compared to chemical synthesis
methods [136].
The essential oil of O. basilicum L. had moderate antibacterial activity against Gram-
negative bacteria. However, combining and formulating the essential oil into chitosan
nanocarriers with nanoencapsulation technology exhibited strong antibacterial and an-
tibiofilm properties against E. coli and S. aureus, resulting in inhibitory zones measuring
15.3 mm and 21 mm, respectively. This combination damages the cell membrane, and there-
fore, it causes the leakage of biological macromolecules [
137
]. Therefore, the combination
has good potential for overcoming Gram-negative resistance against antibiotics. ZnO NP is
one of the nanoparticles, which showed great antibacterial activity against Pseudomonas
aeruginosa, with 20 mm inhibitory zone diameter [138].
ZnO NP synthesized with the O. basilicum L. extract was tested against other bacteria
species and exhibited a great inhibitory zone diameter for S. aureus (19.3 mm), E. coli
(13.2 mm), S. typhimurium (8.2 mm), L. monocytogenes (11.4 mm), B. subtilis (9.3 mm),and
P. aeruginosa (12.4 mm). It also showed great MIC for antibacterial activity, ranging from
0.78
µ
g/mL, 1.56
µ
g/mL, 3.12
µ
g/mL to 6.25
µ
g/mL [
136
]. Along with ZnO NP, copper
oxide nanoparticles (CuO NPs) enhance the antibacterial activity against S. aureus and
E. coli more than the extract itself [
139
]. In addition to the monometallic synthesized
nanoparticle, there are bimetallic synthesized nanoparticles. This is a combination of two
different types of metallic nanoparticles in one particle, which work synergistically [
140
].
In this study, a combination of silver and platinum nanoparticles (AgPt NP) exhibited a
significant inhibitory effect on S. aureus,E. faecalis,E. coli, and K. pneumoniae rather than
the monometallic nanoparticle. The bimetallic particle showed an inhibitory diameter
of
9–25 mm
, whereas the monometallic particle of each nanoparticle only showed an
inhibitory diameter under 10 mm [141].

Plants 2023,12, 4148 17 of 25
Another study demonstrated the green synthesis of reduced graphene oxide (RGO)-
zinc oxide (ZnO) nanocomposite, or RGO-ZnO NCs. It was shown that at a concentration
of 30
µ
g/mL, an inhibition zone was observed for the Cocci strain and E. coli at 20 mm and
10 mm, respectively. RGO-ZnO NCs had antibacterial activity at a small concentration,
whereas the essential oil or extract of O. basilicum L. itself needed higher concentrations to
achieve the same results. This study will also become the basis for the next development
and investigation of RGO-ZnO NCs as potential antioxidant candidates and diabetes treat-
ments [
142
]. Another potential diabetic therapy based on a synthesized silver nanoparticle
was found in O. basilicum L. leaf extract. The result demonstrated inhibitory activity against
α
-amylase—which was higher than antidiabetic medicine acarbose—and high inhibitory
activity against
α
-glucosidase, higher than acarbose and crude extract [
143
]. This finding
suggests the need for alternative therapies for diabetic treatment.
A recent study demonstrated that O. basilicum L. chemical constituents were responsi-
ble for the green biosynthesis of ZnO NPs. In combination with bacterial phages, ZnO NPs
demonstrated antibacterial activity against Salmonella enterica and deformation on biofilm,
which were caused by Staphylococcus sciuri [
144
]. Another study demonstrated the green
synthesis of silver nanoparticles (Ag NPs) in combination with phage ZCSE6 for antibacte-
rial activity against Salmonella enterica. The O. basilicum L. extract works as a bio-reducing
agent in order to create Ag NPs effectively. It was shown that the Ag NPs exhibited an-
tibacterial activity; the minimum concentration to inhibit growth was 6.25
µ
g/mL, and
the minimum bactericidal concentration was 12.5
µ
g/mL. Surprisingly, the Ag NPs in
combination with phage ZCSE6 had great bactericidal activity, with a lower concentration
than the MIC, which suppressed the growth of S. enterica 24 h after treatment [145].
In addition to utilization of the O. basilicum L. extract for synthesizing nanoparticles,
the mucilage from the seed in combination with nanoparticles can create a novel natural
wound dressing. Basil seed mucilage (BSM) was dried and then combined with ZnO
NP to create a hydrogel sponge. As the weight percent (wt%) of ZnO NP increased,
the antibacterial activity of the BSM hydrogel sponge was enhanced. It exhibited great
antibacterial activity at 50 wt% ZnO NP against E. coli and S. aureus, with an inhibitory
zone at 15.9 mm and 16.7 mm, respectively. The increasing ZnO NP wt% content on the
hydrogel sponge also resulted in a slight decrease in thickness, porosity, degree of swelling,
and a slight increase in the water holding capacity. The BSM with ZnO NP is considered
non-toxic to human keratinocyte (HaCat) cells [
146
]. This hydrogel sponge could have the
potential to be commercialized as a natural healthcare product.
In addition to antibacterial functions against human pathogens, the synthesis of silver
nanoparticles (Ag NPs) can also work as a control agent for the management of plant
viral infections. This study tested Ag NPs against cucumber mosaic virus (CMV), which
infects squash. Spraying the foliar containing Ag NPs at a concentration of 100
µ
g/mL
resulted in enhanced growth, delayed indication of disease symptoms, and a significant
reduction of up to 92% in CMV accumulation levels as compared to the non-treated plants.
It also increased the soluble carbohydrate, free radical scavenging activity, antioxidant
enzymes, and total phenolic and flavonoid contents. This finding could be an alternative for
treating plant viral disease instead of using chemical biocides [
147
]. There is a substantial
opportunity in developing a green synthesis of nanoparticles within the O. basilicum L.
extracts, which could be a potential therapy and alternative treatment in many cases of
human diseases.
7.2. Biotechnological Techniques for Improving the Metabolite Production of O. basilicum L.
It is known that the medicinal plant O. basilicum L. is a rich source of valuable phy-
toconstituents [
148
]. The diversity of chemical compounds in O. basilicum L., alone or in
synergy, exhibits some medicinal properties [
149
]. The current production of horticulture
crops is centered on improving the quality, quantity, and safety of products, as well as
yield, in order to meet the demands of the food and health industries, which have a strong
reliance on chemical compounds [
150
]. The advancement of O. basilicum L. production is

Plants 2023,12, 4148 18 of 25
influenced by various aspects, including environmental parameters (light, soil nutrients,
temperature, water, CO
2
levels), cultivars, and cultivation methods [
151
]. Several studies
have demonstrated various experiments on how to improve the chemical compounds
derived from the plant O. basilicum L.
One study showed that narrow-bandwidth light treatments of basil seeds were ob-
served to have relative effects on volatile oils. Light conditions may increase the value
and quality of this herb, which is appreciated for human wellness. Light treatments
could induce the three main compounds in O. basilicum L., which are eugenol, linalool,
and 1,8-cineol (eucalyptol). Eugenol and linalool are induced by blue-red-green (BRG)
light, and 1,8-cineole is induced by BRG, blue-red-yellow (BRY), and blue-red-far-red
(BRFr) light [
152
]. These compounds mainly play a role as antimicrobial and antioxidant
agents [
153
]. The blue and red LED treatments can potentially improve O. basilicum L.
growth and increase the phenolic content of the plants; thus, the different cultivars can also
have a different result. The green cultivar in this study was mostly stimulated by the red
light, and the red cultivar was stimulated by the blue light [154].
In addition to the light treatments, the abiotic (CdCl
2
and AgNO
3
) and biotic (YE)
yeast extract elicitors were found to increase the total amount of phenolic and flavonoid
contents. Chicoric and rosmarinic acid increased with the treatment of CdCl
2
and AgNO
3
at 5
µ
M. Rutin and isoquercetin also increased with the YE treatment, up to 1.6 times and
1.9 times. Meanwhile, the highest amounts of linalool and estragole were observed in the
treatment with AgNO
3
, up to 2.8 times and 0.5 times [
155
]. Arbuscular mycorrhizal fungi
(AMF), which are another type of biotic elicitors, showed a promising capacity in increasing
the production of essential oil, with eugenol and
γ
-cadinene being the compounds with the
highest ratios, which composed the essential oil [156].
Various methods of enhancing the chemical compounds of O. basilicum L. were dis-
cussed above to emphasize the importance of naturally synthesized compounds. One such
approach involves improving the growth factors through light treatments and optimizing
the formulation of biotic or abiotic elicitors.
8. Conclusions
O. basilicum L. is a plant species, which exhibits wide distribution throughout several
regions of the world. Over time, there has been a significant evolution in the understanding
and application of this plant in the context of healthcare. The plant is regarded as a highly
valuable source due to its distinctive chemical composition, which provides a diverse
array of antimicrobial and other medicinal attributes, including anticancer, antioxidant,
antidiabetic, and neuroprotective functions. This plant could change the way in which
drugs are produced, either by isolating pure phytochemical compounds or by combining
several compounds. This could revolutionize the pharmaceutical industry by providing a
natural substitute for synthetic drugs.
The utilization of O. basilicum L. as a medicinal plant has developed over years, start-
ing from community beliefs. The beliefs held by communities were recorded, and many
studies were conducted to prove the efficacy of this plant. Several studies have highlighted
the potential of the aerial parts—particularly the leaves—of this plant for the development
of novel medicines. The phytochemical classes predominantly associated with antimicro-
bial and biomedical activities are polyphenols, terpenes, and phytosterols. Noteworthy
compounds with promising potential for antiviral drug development include carvacrol,
α
-guaiene, ursolic acid, apigenin, stigmasterol, and campesterol. Additionally, compounds
such as linalool, rutin, eugenol, estragole, citral,
α
-pinene, nerolidol, kaempferol, and
trans-
α
-bergamotene could be utilized in the creation of medicines targeting bacteria and
fungi. Moreover, the exploration of new neuroprotective medicines may be facilitated
by novel compounds, such as 5,7-dihydroxy-3
0
,4
0
,5
0
-trimethoxyflavone and 3-hydroxy-
3
0
,4
0
,5
0
-trimethoxyflavone. Furthermore, topical formulation for wound healing has been
demonstrated to be a promising alternative treatment.

Plants 2023,12, 4148 19 of 25
Furthermore, evidence is becoming the key point to be subsequently developed and
formulated into a novel drug. In terms of the plant’s usefulness in the healthcare field,
its distinctive chemical compound, and its higher safety profile, future research should
focus more on formulating this plant into a natural choice in addition to chemical drugs.
As another alternative, this plant could be combined with chemical drugs to create new
efficacy and new mechanisms, which are destined for later commercialization.
Future innovations could come from researching the ways to patent the extraction
methods, formulating a standardized approach to boost the chemical compound contents,
conducting more clinical research in a mechanistic and molecular way, and advancing it
up to the industrial stage. This will allow not only gaining a deeper understanding of the
mechanistic action, but it will also lead the path to developing more effective, safer drugs
and reduce undesirable side effects.
Author Contributions:
Conceptualization and writing—original draft preparation, N.S.A.; writ-
ing—review and editing, and supervision, B.I., M.M., J.K., D.O. and F.D.; writing—review and
editing, and drawing chemical constituents, W.S. and K.F. All authors have read and agreed to the
published version of the manuscript.
Funding:
This work was funded by Universitas Padjadjaran through the Hibah Riset Percepatan
Lektor Kepala awarded to Mia Miranti under grant number 159/UN6.3.1/PT.00/2023.
Data Availability Statement: This study did not report any data.
Acknowledgments:
The authors would like to thank Karina Kalasuba and Sulistya Ika Akbari for
their assistance during the trip.
Conflicts of Interest: The authors declare no conflict of interest.
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