High level extracellular production of a truncated alkaline beta-mannanase from alkaliphilic Bacillus sp N16-5 in Escherichia coli by the optimization of induction condition and fed-batch fermentation

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	High level extracellular production of a truncated alkaline beta-mannanase from alkaliphilic Bacillus sp N16-5 in Escherichia coli by the optimization of induction condition and fed-batch fermentation
	Zheng, Hongchen 1,2; Yu, Zhenxiao 1,2; Fu, Xiaoping 1,2; Li, Shufang 1,2; Xu, Jianyong 1,2; Song, Hui 1,2; Ma, Yanhe 1
刊名	JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
	2016-07-01
卷号	43 期号:7 页码:977-987
关键词	Alkaline beta-mannanase Truncation Escherichia coli Triton X-100 One-time feeding
英文摘要	To improve the extracellular production of alkaline beta-mannanase from alkaliphilic Bacillus sp. N16-5 in Escherichia coli, two truncated recombinant mannanases (32a-ManAR2 and 22b-ManAR2) were obtained. Compared with the full-length mannanases (32a-ManAR1 and 22b-ManAR1), the truncated mannanases not only showed higher secretion rate, but also exhibited higher thermostability and alkalistability. The K-m value (11 mg/mL) of 32a-ManAR2 was higher than that (1.46 mg/mL) of 32a-ManAR1. The specific activity of 22b-ManAR2 was 2.7 times higher than that of 22b-ManAR1. 22b-ManAR2 showed the highest k(cat)/K-m value of 602.7 ml/mg s. The parameters of induction for recombinant mannanase production of E. coli BL21 (pET32a-manAR2) and E. coli BL21 (pET22b-manAR2) were subsequently optimized. The yield of soluble mannanase was found to be enhanced with lower induction temperature (25 degrees C), lower IPTG concentration (0.01-0.05 mM), and Triton X-100 supplement (0.1 %) in a shake flask. Moreover, a one-time feeding strategy and Triton X-100 supplement were applied in production of 22b-ManAR2 in a 10 L fermentor. The productivity of the total soluble mannanase reached 9284.64 U/mL with the extracellular rate of 74 % at 46 h of fermentation, which was the highest productive level of alkaline beta-mannanase in recombinant E. coli to date.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biotechnology & Applied Microbiology
研究领域[WOS]	Biotechnology & Applied Microbiology
关键词[WOS]	PICHIA-PASTORIS ; GENE CLONING ; EXPRESSION ; PULLULANASE ; STRATEGY
收录类别	SCI
语种	英语
WOS记录号	WOS:000379625600008
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/2912]
专题	天津工业生物技术研究所_基因工程与微生物应用技术研究组宋恢_期刊论文
作者单位	1.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Ind Enzymes Natl Engn Lab, Tianjin 300308, Peoples R China 2.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Tianjin Key Lab Ind Biol Syst & Bioproc Engn, 32 XiQiDao,Tianjin Airport Econ Pk, Tianjin 300308, Peoples R China
推荐引用方式 GB/T 7714	Zheng, Hongchen,Yu, Zhenxiao,Fu, Xiaoping,et al. High level extracellular production of a truncated alkaline beta-mannanase from alkaliphilic Bacillus sp N16-5 in Escherichia coli by the optimization of induction condition and fed-batch fermentation[J]. JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY,2016,43(7):977-987.
APA	Zheng, Hongchen.,Yu, Zhenxiao.,Fu, Xiaoping.,Li, Shufang.,Xu, Jianyong.,...&Ma, Yanhe.(2016).High level extracellular production of a truncated alkaline beta-mannanase from alkaliphilic Bacillus sp N16-5 in Escherichia coli by the optimization of induction condition and fed-batch fermentation.JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY,43(7),977-987.
MLA	Zheng, Hongchen,et al."High level extracellular production of a truncated alkaline beta-mannanase from alkaliphilic Bacillus sp N16-5 in Escherichia coli by the optimization of induction condition and fed-batch fermentation".JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY 43.7(2016):977-987.