Using inexpensive substrate to achieve high-level lipase A secretion by Bacillus subtilis through signal peptide and promoter screening
Using inexpensive substrate to achieve high-level lipase A secretion by Bacillus subtilis through signal peptide and promoter screening
复制标题
利用廉价底物通过信号肽和启动子筛选实现枯草芽孢杆菌高水平脂肪酶A分泌
DOI：
10.1016/j.procbio.2020.08.010
复制
发表时间：
2020-12-01
影响因子：
4.4
通讯作者：
Zhu, Mingjun
中科院分区：
文献类型：
--
作者：
Wu, Fengyi;Ma, Jiayuan;Zhu, Mingjun
脂肪酶A因其在食品造纸工业、医药化工和日用化学品中的应用而显示出巨大的商业价值。在这项研究中，我们首先对不同的信号肽和启动子进行了比较，以构建脂肪酶A高产菌株。枯草芽孢杆菌L25在37℃下发酵12h，最大胞外脂肪酶活力为64.9U/mL，其中脂肪酶基因由SPlipA引导，由P-43-P-HAG调控。另一个目的是确定是否在培养基中添加甘油和链霉菌的混合物。SCut-3消化鸡毛水解液有利于脂肪酶的产生。设计并进行了系统的优化实验。最终在C/N比为0.5时，在特级肉汤培养基中获得了较高的脂肪酶活力(1164.9 U/mL)。本工作论证了枯草杆菌L25以甘油和微生物处理的鸡毛水解液为廉价碳源和氮源工业化生产脂肪酶A的可行性。
Lipase A exhibits great commercial value due to its applications in the food and paper industry, pharmaceutical chemistry and household chemicals. In this study, we first conducted a comparison of different signal peptides and promoters for the construction of the productive lipase A strain. The maximum extracellular lipase activity was identified as 64.9 U/mL after 12-h fermentation at 37 degrees C by B. subtilis L25 in which the lipase gene was led by SPlipA and controlled by P-43-P-hag. Another aim was to determine whether supplementing a culture medium with a combination of glycerol and Streptomyces sp. SCUT-3 digested chicken feather hydrolysates was beneficial to lipase production. Systematic optimization experiments were designed and carried out. Finally, a satisfactory lipase level of 1164.9 U/mL was accomplished in Terrific-Broth medium at a C/N ratio of 0.5 (v/v). This work demonstrates the feasibility of B. subtilis L25 for lipase A industrial production using glycerol and microbes treated chicken feather hydrolysates as inexpensive carbon and nitrogen source.