Overexpression of an endogenous raw starch digesting mesophilic α-amylase gene in Bacillus amyloliquefaciens Z3 by in vitro methylation protocol.
Overexpression of an endogenous raw starch digesting mesophilic α-amylase gene in Bacillus amyloliquefaciens Z3 by in vitro methylation protocol.
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通过体外甲基化协议在解淀粉芽孢杆菌 Z3 中过度表达内源性生淀粉消化嗜温 α-淀粉酶基因。
DOI：
10.1002/jsfa.10332
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发表时间：
2020-02
影响因子：
4.1
通讯作者：
Hongbo Zhou
中科院分区：
文献类型：
--
作者：
Shizhe Tang;Tingliang Xu;Jing Peng;Kaiyan Zhou;Yuling Zhu;Wenbo Zhou;Haina Cheng;Hongbo Zhou
背景：乳酸α-淀粉酶在低温下以高催化速率有效地发挥作用，并且淀粉水解需要较少的能量。解淀粉芽孢杆菌（Bacillus amyloliquefaciens）是嗜中温α-淀粉酶的重要生产者。然而，由于限制修饰系统（RMS）的存在，将外源DNA导入野生型B。解淀粉菌特别棘手。结果：α-淀粉酶产生菌B。筛选到一株解淀粉菌Z3，并将其作为内源α-淀粉酶基因表达的宿主。在重组质粒pWB 980-amyZ 3中进行体外甲基化。体外甲基化后，转化效率可达0.96 × 102 CFU/μ g质粒DNA。选择α-淀粉酶分泌能力最高的阳性菌株BAZ 3 -16进行进一步的实验。在摇瓶和5L搅拌罐中，BAZ 3 -16菌株的α-淀粉酶活力分别为288.70 ± 16.15 U mL-1和386.03 ± 16.25 U mL-1。B。Z3表达系统具有良好的遗传稳定性和高水平的目的蛋白胞外产量。此外，在水解生淀粉的过程中，确定了AmyZ 3与淀粉葡萄糖苷酶的协同相互作用。100 g L-1的玉米原淀粉和100 g L-1的木薯原淀粉在24 h的水解度分别达到92.34 ± 3.41%和81.30 ± 2.92%。结论：质粒DNA的甲基化消除了转化B的实质性障碍。解淀粉杆菌菌株Z3。此外，α-淀粉酶AmyZ 3和菌株BAZ 3 -16具有优异的淀粉水解能力，在淀粉工业中具有重要的应用价值。本文受版权保护。All rights reserved.
BACKGROUND.Mesophilic α-amylases function effectively at low temperatures with high rates of catalysis and require less energy for starch hydrolysis. Bacillus amyloliquefaciens is an essential producer of mesophilic α-amylases. However, due to the existence of the restriction-modification system (RMS), introducing exogenous DNAs into wild-type B. amyloliquefaciens is especially tricky..RESULTS.α-Amylase producer B. amyloliquefaciens strain Z3 was screened and used as host for endogenous α-amylase gene expression. In vitro methylation was performed in recombinant plasmid pWB980-amyZ3. With the in vitro methylation, the transformation efficiency was increased to 0.96 × 102 CFUs per microgram plasmid DNA. A positive transformant BAZ3-16 with the highest α-amylase secreting capacity was chosen for further experiments. The α-amylase activity of strain BAZ3-16 reached 288.70 ± 16.15 U mL-1 in flask and 386.03 ± 16.25 U mL-1 in 5-L stirred-tank fermenter, respectively. The B. amyloliquefaciens Z3 expression system shows excellent genetic stability and high-level extracellular production of the target protein. Moreover, the synergistic interaction of AmyZ3 with amyloglucosidase was determined during the hydrolysis of raw starch. The hydrolysis degree reached 92.34 ± 3.41% for 100 g L-1 raw corn starch and 81.30 ± 2.92% for 100 g L-1 raw cassava starch after 24 h, respectively..CONCLUSION.Methylation of the plasmid DNA removes a substantial barrier for transformation of B. amyloliquefaciens strain Z3. Besides, exceptional ability to hydrolyze starch makes α-amylase AmyZ3 and strain BAZ3-16 valuable in the starch industry. This article is protected by copyright. All rights reserved.
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影响因子：
2.9
作者：
S. Özdemir;Fatma Matpan;K. Güven;Zübeyde Baysal
通讯作者：
S. Özdemir;Fatma Matpan;K. Güven;Zübeyde Baysal
DOI：
10.1007/bf00264941
发表时间：
1977-03
期刊：
Molecular and General Genetics MGG
影响因子：
--
作者：
T. Uozumi;T. Hoshino;K. Miwa;S. Horinouchi;T. Beppu;K. Arima
通讯作者：
T. Uozumi;T. Hoshino;K. Miwa;S. Horinouchi;T. Beppu;K. Arima
解淀粉芽孢杆菌和枯草芽孢杆菌的比较基因组分析揭示了适应植物相关生境的进化特征
影响因子：
5.2
作者：
Zhang N;Yang D;Kendall JR;Borriss R;Druzhinina IS;Kubicek CP;Shen Q;Zhang R
通讯作者：
Zhang R
影响因子：
6.3
作者：
Viktor MJ;Rose SH;van Zyl WH;Viljoen-Bloom M
通讯作者：
Viljoen-Bloom M
影响因子：
2.6
作者：
Lim, WJ;Park, SR;Yun, HD
通讯作者：
Yun, HD