| CHAN Zhuhua,CHEN Xinglin,HOU Yanping,GAO Boliang,ZHAO Chungui,YANG Suping,ZENG Runying. 2018. Enhanced a novel β-agarase production in recombinant Escherichia coli BL21 (DE3) through induction mode optimization and glycerol feeding strategy. Acta Oceanologica Sinica, 37(2):110－118 |
| Enhanced a novel β-agarase production in recombinant Escherichia coli BL21 (DE3) through induction mode optimization and glycerol feeding strategy |
| 一种诱导模式和甘油补料优化策略提高重组大肠杆菌BL21（DE3）β-琼胶酶的产量 |
| Received:December 28, 2016 Revised:September 21, 2011 |
| DOI：10.1007/s13131-018-1172-x |
| Key words:β-agarase Escherichia coli process optimization glycerol feeding strategy neoagarooligosaccharide |
| 中文关键词: β-琼胶酶 大肠杆菌 过程优化 甘油补料策略 新琼寡糖 |
| 基金项目: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. |
| Author Name | Affiliation | E-mail | | CHAN Zhuhua | Department of Bioengineering and Biotechnology, Huaqiao University, Xiamen 361021, China
State Key Laboratory Breeding Base of Marine Genetic Resource, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China | | | CHEN Xinglin | State Key Laboratory Breeding Base of Marine Genetic Resource, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China | | | HOU Yanping | State Key Laboratory Breeding Base of Marine Genetic Resource, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China | | | GAO Boliang | State Key Laboratory Breeding Base of Marine Genetic Resource, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China
School of Life Sciences, Xiamen University, Xiamen 361102, China | | | ZHAO Chungui | Department of Bioengineering and Biotechnology, Huaqiao University, Xiamen 361021, China | | | YANG Suping | Department of Bioengineering and Biotechnology, Huaqiao University, Xiamen 361021, China | yangsuping@hqu.edu.cn | | ZENG Runying | State Key Laboratory Breeding Base of Marine Genetic Resource, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China | |
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| Abstract: |
| 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. |
| 中文摘要: |
| 琼胶酶是作用于琼胶的水解酶，在食品、化妆品和制药工业中有广泛的应用。本文建立了一种基于诱导模式和甘油补料优化的高细胞密度和高产β-琼胶酶策略，同时可以较好的控制乙酸盐产量。首先，在诱导前期采用不同的比生长速率（μ）的甘油指数补料策略。结果表明，低的比生长速率（μ=0.2）是细胞生长和β-琼胶酶产生的最佳条件。其次，研究了诱导阶段诱导温度和诱导物浓度对细胞生长和β-琼胶酶产生的影响。当异丙基-β-d-硫代半乳糖苷（IPTG）诱导时，在20℃下0.8mmol/L的IPTG诱导策略对β-琼胶酶的产生效果最佳。采用1.0g/（L·h）的乳糖连续补料策略诱导培养，β-琼胶酶活性达到112.5U/mL，是目前报道的产量最高的β-琼胶酶。此外，β-琼胶酶能直接酶解龙须菜粉，产生新琼寡糖，水解产物为新琼胶四糖（NA4）和新琼胶六糖（NA6）。本文的研究为β-琼胶酶的工业化生产和应用提供了较好的理论依据。 |
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