Identification of yeast strains isolated from marcha in Sikkim, a microbial starter for amylolytic fermentation

Experimental Farm, Shinshu University, Shonai, Nagano, Japan
International Journal of Food Microbiology (Impact Factor: 3.16). 04/2005; 99(2):135-46. DOI: 10.1016/j.ijfoodmicro.2004.08.011
Source: PubMed

ABSTRACT Marcha or murcha is a traditional amylolytic starter used to produce sweet-sour alcoholic drinks, commonly called jaanr in the Himalayan regions of India, Nepal, Bhutan, and Tibet (China). The aim of this study was to examine the microflora of marcha collected from Sikkim in India, focusing on yeast flora and their roles. Twenty yeast strains were isolated from six samples of marcha and identified by genetic and phenotypic methods. They were first classified into four groups (Group I, II, III, and IV) based on physiological features using an API test. Phylogenetic, morphological, and physiological characterization identified the isolates as Saccharomyces bayanus (Group I); Candida glabrata (Group II); Pichia anomala (Group III); and Saccharomycopsis fibuligera, Saccharomycopsis capsularis, and Pichia burtonii (Group IV). Among them, the Group I, II, and III strains produced ethanol. The isolates of Group IV had high amylolytic activity. Because all marcha samples tested contained both starch degraders and ethanol producers, it was hypothesized that all four groups of yeast (Group I, II, III, and IV) contribute to starch-based alcohol fermentation.

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    • "The most well-known and commercially significant yeasts that been primarily used for bioethanol production are the related species and strains of Saccharomyces cerevisiae [4]. These organisms have long been utilized to ferment the sugars of rice, wheat, barley, and corn to produce alcoholic beverages and in the backing industry [5]. One yeast cell can ferment approximately its own weight of glucose per hour. "
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    • "S. fibuligera, the most abundant yeast species isolated in this study, can secrete a large amount of a-amylase, glucoamylase, acid proteases and b-glucosidase, and had been applied in the fermentation industry (Chi et al., 2009). It had been reported that S. fibuligera was the principal amylolytic microorganism in traditional alcohol starters such as Loog-pang, Marcha and banh men (Limtong, Sintara, Suwannarit, & Lotong, 2002; Thanh, Mai, & Tuan, 2008; Tsuyoshi et al., 2005). S. cerevisiae is the most effective ethanol producer known so far (Vaughan-Martini & Martini, 1995), and has been reported to be the commonest yeast in indigenous fermented foods and beverages, where it has been shown to be very important, especially in the fermentation of cereals and alcoholic beverages (Jespersen, 2003). "
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    ABSTRACT: Hong Qu glutinous rice wine, one of the most popular traditional rice wines in China, is brewed from glutinous rice with the addition of two traditional fermentation starters --- Hong Qu (hóng qū) and Yao Qu (yào qū). The aim of this study was to investigate the yeast flora present in these fermentation starters using a combination of culture-dependent and -independent molecular biological methods. Molecular identification of a total of 500 yeast isolate from 10 kinds of representative wine fermentation starters (5 Hong Qu and 5 Yao Qu) was achieved by a combination of polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) analysis of the internal transcribed spacer region ITS1-5.8S-ITS2 and sequencing of the D1/D2 domain of the 26S rRNA gene. The PCR-RFLP method generated 13 different ITS/RFLP profiles. Sequencing identification allowed identifying 13 different ITS/RFLP profiles into 12 different species of yeast belong to eight different genera (Pichia, Saccharomyces, Candida, Cryptococcus, Rhodotorula, Sporobolomyces, Rhodosporidium and Saccharomycopsis). Species diversity was thus considerable: Saccharomycopsis fibuligera was the most commonly isolated specie, followed by Saccharomyces cerevisiae. On the other hand, the yeast diversity associated with wine starters for Hong Qu glutinous rice wine was also investigated through culture-independent method using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) patterns and sequencing of the DNA bands. The results revealed by culture-independent method were almost the same as that of culture-dependent methodology. However, several yeast species can not be detected by traditional microbiological procedures but can be detected using PCR-DGGE analysis. Similarly, some species could only be detected by culture-dependent method. For example, Sporobolomyces nylandii was detected existing in wine starter samples (Q7 and Q10) through culture-dependent method, but not detected in wine starters for Hong Qu glutinous rice wine through fungal PCR-DGGE assay. Therefore, it is recommended that polyphonic approaches based on culture-dependent and culture independent methods seem the best strategy to get a more comprehensive picture of the microbial diversity in traditional wine starters. The results enrich our knowledge of rice wine-related yeasts, and can be used to promote the development of the traditional brewing industry.
    Food Control 05/2013; 34(1):183-190. DOI:10.1016/j.foodcont.2013.04.020 · 2.82 Impact Factor
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    • "Ethnic people of the Himalayan regions of India including North East regions, Nepal, Bhutan and China domesticate consortium of native microorganisms consisting of amylolytic filamentous molds and alcohol-producing yeasts in the form of dry, flattened or round cake-like starter locally called hamei in Manipur state of India (Jeyaram et al. 2008), marcha in Sikkim and Darjeeling hills in India, Nepal and Bhutan (Tsuyoshi et al. 2005), which are similar to ragi and brem of Indonesia, bubod of Philippines, chu of China, naruk of Korea, men of Vietnam and loogpang of Thailand (Sujaya et al. 2004; Tamang 2010). Marcha is traditionally used as starter to produce various alcoholic beverages, such as kodo ko jaanr from finger millets (Thapa and Tamang 2004) and bhaati jaanr from rice (Tamang and Thapa 2006). "
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    ABSTRACT: Autochthonous strains of Saccharomyces cerevisiae from traditional starters used for the production of rice-based ethnic fermented beverage in North East India were examined for their genetic polymorphism using mitochondrial DNA-RFLP and electrophoretic karyotyping. Mitochondrial DNA-RFLP analysis of S. cerevisiae strains with similar technological origins from hamei starter of Manipur and marcha starter of Sikkim revealed widely separated clusters based on their geographical origin. Electrophoretic karyotyping showed high polymorphism amongst the hamei strains within similar mitochondrial DNA-RFLP cluster and one unique karyotype of marcha strain was widely distributed in the Sikkim-Himalayan region. We conceptualized the possibility of separate domestication events for hamei strains in Manipur (located in the Indo-Burma biodiversity hotspot) and marcha strains in Sikkim (located in Himalayan biodiversity hotspot), as a consequence of less homogeneity in the genomic structure between these two groups, their clear separation being based on geographical origin, but not on technological origin and low strain level diversity within each group. The molecular markers developed based on HinfI-mtDNA-RFLP profile and the chromosomal doublets in chromosome VIII position of Sikkim-Himalayan strains could be effectively used as geographical markers for authenticating the above starter strains and differentiating them from other commercial strains.
    Antonie van Leeuwenhoek 06/2011; 100(4):569-78. DOI:10.1007/s10482-011-9612-z · 2.14 Impact Factor
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