Cloning and Bioinformatical Analysis of α-acetolactate Decarboxylase Gene (alsD) in Bacillus subtilis

doi:10.11924/j.issn.1000-6850.casb18020058

Abstract

Abstract: α-acetolactate decarboxylase is one of the key enzymes in 2,3-butanediol biosynthesis pathway, and the alsD gene is analyzed by bioinformatics method. Primers are designed based on the gene sequence of α- acetolactate decarboxylase gene (alsD) in GenBank. Bacillus subtilis CICC10026 gene fragment alsD is amplified by PCR which the template is Bacillus subtilis CICC10026 genomic DNA. The fragment is sequenced and analyzed by bioinformatics software for homology analysis, secondary structure and functional analysis and 3D structure prediction. The results showed that the obtained alsD fragment was 768 bp without base mutation, and the fragment was the alsD gene sequence of Bacillus subtilis CICC10026. The results of this study lay a foundation for the construction of engineering strains which could produce 2,3-butanediol.

References

[1] Kim S J, Sim H J, Kim J W, et al. Enhanced production of 2,3-butanediol from xylose by combinatorial engineering of xylose metabolic pathway and cofactor regeneration in pyruvate decarboxylase-deficient Saccharomyces cerevisiae[J]. Bioresource Technology, 2017, 245: 1551-1557.
[2] Jooa J, Leeb S, Yoo H, et al. Improved fermentation of lignocellulosic hydrolysates to 2,3-butanediol through investigation of effects of inhibitory compounds by Enterobacter aerogenes[J]. Chemical Engineering Journal, 2016(306), 916-924.
[3] 宋源泉, 许赟珍, 李强, 等. 2,3-丁二醇的发酵生产[J]. 化工进展, 2011, 30(5): 1069-1077.
[4] 杨世哲, 朱冬梅. 甲乙酮的合成及其应用的研究进展[J]. 广州化工, 2011 (8), 15-17.
[5] 刘德龙, 张玉苍, 何连芳, 等. 生物转化法生产2,3-丁二醇的研究进展[J]. 食品工程, 2011, 3: 143-146.
[6] Guo X W, Wang Y Z, Guo J, et al. Efficient production of 2,3-butanediol from cheese whey powder (CWP) solution by Klebsiella pneumoniae through integrating pulsed fed-batch fermentation with a two-stage pH control strategy [J]. Bioresource Technology, 2017, 245: 1386-1392.
[7] Guo X W, Zhang Y H, Cao C H, et al. Enhanced production of 2,3-butanediol by overexpressing acetolactate synthase and acetoin reductase in Klebsiella pneumoniae[J]. Biotechnology and Applied Biochemistry, 2014, 61(6): 707-715.
[8] Park J H, Choi MA, Kim YJ, et al. Engineering of Klebsiella oxytoca for production of 2,3-butanediol via simultaneous utilization of sugars from a Golenkinia sp. hydrolysate [J]. Fuel, 2017, 203: 469-477.
[9] Kim D K, Rathnasingh C, Song H, et al. Metabolic engineering of a novel Klebsiella oxytoca strain for enhanced 2,3-butanediol production[J]. Journal of Bioscience and Bioengineering, 2013, 116(2): 186-192.
[10] Wang Q, Chen T, Zhao X, et al. Metabolic engineering of thermophilic Bacillus licheniformis for chiral pure D-2,3-butanediol production. Biotechnology and Bioengineering, 2012(109), 1610–1621.
[11] Ma K, He M, You H, et al. Improvement of (R,R)-2,3-butanediol production from corn stover hydrolysate by cell recycling continuous fermentation[J]. Chemical Engineering Journal, 2018, 15(1): 361-369.
[12] Lee S, Kim C, Choi H, et al. Process strategy for 2,3-butanediol production in fed-batch culture by acetate addition [J]. Journal of Industrial and Engineering Chemistry, 2017, 25(12): 157-162.
[13] Ji X J, Huang H, Yang P K. Microbial 2,3-butanediol production: a state-of-the-art review[J]. Biotechnology Advances, 2011, 29(3): 351-364.
[14] 付晶, 王萌, 刘维喜. 生物法制备2,3-丁二醇的最新进展[J]. 化学进展, 2012, 11(24): 2268-2276.
[15] Ryosuke Y, Kazuki W, Ryosuke M, et al. Efficient production of 2,3-butanediol by recombinant Saccharomyces cerevisiae through modulation of gene expression by cocktail d-integration [J]. Bioresource Technology, 2017, 2445: 1558-1566.
[16] Kim S, Hahn JS. Efficient production of 2,3-butanediol in Saccharomyces cerevisiae by eliminating ethanol and glycerol production and redox rebalancing[J]. Metabolic Engineering, 2015, 31: 94-101.
[17] Ng C Y, Jung M Y, Lee J, et al. Production of 2,3-butanediol in Saccharomyces cerevisiae by in silico aided metabolic engineering [J]. Microbial Cell Factories, 2012, 11: 68.
[18] Kim S J, Seo S O, Jin Y S, et al. Production of 2,3-butanediol by engineered Saccharomyces cerevisiae [J]. Bioresource Technology, 2013, 146: 274-281.
[19] Choi M, Kim S J, Kim J W, et al. Molecular cloning and express of Enterobacter aerogees α-acetolactate decarboxylase in pyruvate decarboxylase-deficient Saccharomyces cerevisiae for efficient 2,3-butanediol production[J]. Process Biochemistry, 2016, 51(2): 170-176.
[20] Ji X J, Huang H, Ouyang PK. Microbial 2,3-butanediol production: a state-of-the-art review[J]. Biotechnology Advances, 2011, 29(3): 351-364.
[21] 王青艳, 谢能中, 黎贞崇, 等. 微生物法合成(R,R)-2,3-丁二醇的研究进展与展望[J]. 基因组学与应用生物学, 2014 , 33(6), 1367-1373.
[22] Zhang L J, Ran C, Zhao H. Metabolic engineering of a Saccharomyces cerevisiae strain capable of simultaneously utilizing glucose and galactose to produce enantiopure (2R,3R)-butanediol[J]. Metabolic Engineering, 2014, 23(5): 92-99.
[23] 黄守锋, 裴芳艺, 王长丽, 等. 利用酿酒酵母工程菌株生产2,3-丁二醇的研究进展[J]. 食品安全质量检测学报, 2015, 6(10), 3928-3934.
[24] 王长丽, 孙雯, 黄守锋, 等. 产酸克雷伯氏菌(Klebsiella oxytoca)α-乙酰乳酸脱羧酶基因(BudA)的克隆及生物信息学分析[J]. 黑龙江大学自然科学学报, 2016, 127(10), 795-801.
[25] 李静静. 高产α-乙酰乳酸脱羧酶重组枯草芽孢杆菌的构建及其发酵优化[D]. 无锡: 江南大学, 2013.
[26] 黄广庆. 缺失GPD2基因对工业酿酒酵母乙醇发酵的影响[D]. 广州: 暨南大学, 2013.