Genome sequence of Bacillus licheniformis WX-02.
ABSTRACT Bacillus licheniformis is an important bacterium that has been used extensively for large-scale industrial production of exoenzymes and peptide antibiotics. B. licheniformis WX-02 produces poly-gamma-glutamate increasingly when fermented under stress conditions. Here its genome sequence (4,270,104 bp, with G+C content of 46.06%), which comprises a circular chromosome, is announced.
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ABSTRACT: Poly-γ-glutamate (PGA), a novel polyamide material with industrial applications, possesses a nylon-like backbone, is structurally similar to polyacrylic acid, is biodegradable and is safe for human consumption. PGA is frequently found in the mucilage of natto, a Japanese traditional fermented food. To date, three different types of PGA, namely a homo polymer of d-glutamate (D-PGA), a homo polymer of l-glutamate (L-PGA), and a random copolymer consisting of d- and l-glutamate (DL-PGA), are known. This review will detail the occurrence and physiology of PGA. The proposed reaction mechanism of PGA synthesis including its localization and the structure of the involved enzyme, PGA synthetase, are described. The occurrence of multiple carboxyl residues in PGA likely plays a role in its relative unsuitability for the development of bio-nylon plastics and thus, establishment of an efficient PGA-reforming strategy is of great importance. Aside from the potential applications of PGA proposed to date, a new technique for chemical transformation of PGA is also discussed. Finally, some techniques for PGA and its derivatives in advanced material technology are presented.Microbial Biotechnology 07/2013; · 3.21 Impact Factor
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ABSTRACT: D-2,3-butanediol has many industrial applications such as chiral reagents, solvents, anti-freeze agents, and low freezing point fuels. Traditional D-2,3-butanediol producing microorganisms, such as Klebsiella pneumonia and K. xoytoca, are pathogenic and not capable of producing D-2,3-butanediol at high optical purity. Bacillus licheniformis is a potential 2,3-butanediol producer but the wild type strain (WX-02) produces a mix of D- and meso-type isomers. BudC in B. licheniformis is annotated as 2,3-butanediol dehydrogenase or acetoin reductase, but no pervious experiment was performed to verify this hypothesis. We developed a genetically modified strain of B. licheniformis (WX-02 DeltabudC) as a D-2,3-butanediol producer with high optimal purity. A marker-less gene deletion protocol based on a temperature sensitive knock-out plasmid T2-Ori was used to knock out the budC gene in B. licheniformis WX-02. The budC knock-out strain successfully abolished meso-2,3-butanediol production with enhanced D-2,3-butanediol production. No meso-BDH activity was detectable in cells of this strain. On the other hand, the complementary strain restored the characteristics of wild strain, and produced meso-2,3-butanediol and possessed meso-BDH activity. All of these data suggested that budC encoded the major meso-BDH catalyzing the reversible reaction from acetoin to meso-2,3-butanediol in B. licheniformis. The budC knock-out strain produced D-2,3-butanediol isomer only with a high yield of 30.76 g/L and a productivity of 1.28 g/L-h. We confirmed the hypothesis that budC gene is responsible to reversibly transfer acetoin to meso-2,3-butanediol in B. licheniformis. A mutant strain of B. licheniformis with depleted budC gene was successfully developed and produced high level of the D-2,3- butanediol with high optimal purity.Biotechnology for Biofuels 01/2014; 7(1):16. · 5.55 Impact Factor
Genome Sequence of Bacillus licheniformis WX-02
Wuming Yangtse, Yinhua Zhou, Yang Lei, Yimin Qiu, Xuetuan Wei, Zhixia Ji, Gaofu Qi, Yangchun Yong, Lingling Chen,
and Shouwen Chen
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, People’s Republic of China
Bacillus licheniformis is an important bacterium that has been used extensively for large-scale industrial production of exoen-
zymes and peptide antibiotics. B. licheniformis WX-02 produces poly-gamma-glutamate increasingly when fermented under
is used to produce enzymatic agents (12), peptide antibiotics (7),
acetoin (8), and poly-?-glutamic acid (?-PGA) (1, 2), etc. B. li-
cheniformis WX-02 was isolated from saline soil in Yingcheng
produce larger amounts of ?-PGA under stress conditions than
under normal ones; the stresses refer to high salt, high tempera-
ture, caustic alkali, ultrasonic shock, and so on. The yield of
?-PGA can be raised 5.28 times against that of the control group
when the salt concentration reaches 8% (13). In order to explore
the mechanism of high production of ?-PGA under adverse cir-
licheniformis WX-02 has been maintained at the China Center for
Type Culture Collection and designated B. licheniformis CCTCC
Whole-genome sequencing was performed using a Solexa
genome analyzer (BGI; Shenzhen, China) using a shotgun
strategy, which produced paired reads totaling ?555 Mb with
were processed and assembled into 45 contigs and 29 scaffolds
(5). Gaps between contigs were closed by combinatorial PCR
and sequencing amplicons by primer walking. Finally, this as-
sembling process produced 3 large scaffolds in one potential
circular chromosome. Open reading frames were identified using
&group?programs&subgroup?gfindb), Prodigal (http://compbio
.ornl.gov/prodigal/), and Glimmer (http://www.ncbi.nlm.nih.gov
/genomes/MICROBES/glimmer_3.cgi) and BLAST against the Ref-
Seq database. tRNA and rRNA genes were identified by the
tRNAScan and RNAmmer software programs, respectively (4, 6).
The genome of strain CCTCC M208065 is 4,270,104 bp long with a
GC content of 46.06%, containing 4,320 open reading frames, 72
M208065 shares great similarity (94.8%) with that of ATCC 14580,
about 76 kb in this genome sharing very low homology with the
ment encodes the DNA replication initiator protein, conjugation
protein, RNA polymerase sigma factors, transcriptional regulators,
acillus licheniformis is a Gram-positive bacterium which in-
in regulation of ?-PGA synthesis (3, 11). The comP gene in the
ATCC 14580 strain is inactivated by two insertional fragments,
the comP gene between these two strains might be associated with
idence is needed to verify this.
paper is the first version, AHIF01000000.
(grant no. 30970036 and no. 31170046).
We are grateful to Sun Ming for his kind and generous suggestions.
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Received 5 April 2012 Accepted 10 April 2012
Address correspondence to Shouwen Chen, email@example.com.
Copyright © 2012, American Society for Microbiology. All Rights Reserved.
July 2012 Volume 194 Number 13Journal of Bacteriologyp. 3561–3562jb.asm.org
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