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Enhanced a novel β-agarase production in recombinant Escherichia coli BL21(DE3) through induction mode optimization and glycerol feeding strategy

查看全文 作  者:CHAN [1,2]Zhuhua;CHEN [2]Xinglin;HOU [2]Yanping;GAO [2,3]Boliang;ZHAO [1]Chungui;YANG [1]Suping;ZENG [2]Runying 高影响力作者 机构地区:[1]Department of Bioengineering and Biotechnology, Huaqiao University;[2]State Key Laboratory Breeding Base of Marine Genetic Resource, Third Institute of Oceanography State Oceanic Administration;[3]School of Life Sciences, Xiamen University高影响力机构 出  处:《Acta Oceanologica Sinica》索引2018年第37卷第2期,共9页高影响力期刊 基  金:The Public Science and Technology Research Funds Projects of Ocean under contract No.201505026;the Fujian Province Natural Science Foundation under contract Nos 2016J01160 and 2017N0015;the Scientific Research Foundation of Third Institute of Oceanography,SOA under contract No.2016038 摘  要:Agarases are hydrolytic enzymes that act on the hydrolysis of agar and have a broad range of applications in food,cosmetics and pharmaceutical industries. In this study, a glycerol feeding strategy based on induction mode optimization for high cell density and β-agarase production was established, which could effectively control acetate yield. First, exponential feeding strategy of glycerol with different overall specific growth rates(μ) was applied in the pre-induction phase. The results showed that the low μ(μ=0.2) was suggested to be the optimal for cell growth and β-agarase production. Second, the effects of induction temperature and the inducer concentration on cell growth and β-agarase production were investigated in the post-induction phase. When induced by isopropyl-β-d-thiogalactoside(IPTG), the strategy of 0.8 mmol/L IPTG induction at 20℃ was found to be optimal for β-agarase production. When cultivation was induced by continuous lactose feeding strategy of 1.0 g/(L·h), the β-agarase activity reached 112.5 U/mL, which represented the highest β-agarase production to date.Furthermore, the β-agarase was capable of degrading G. lemaneiformis powder directly to produce neoagarooligosaccharide, and the hydrolysates were neoagarotetraose(NA4) and neoagarohexaose(NA6). The overall research may be useful for the industrial production and application of β-agarase. 关 键 词:工业生产 模式优化 甘油 细胞生长 水解作用 IPTG 化妆品 盐产量
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