融合短肽促进碱性果胶酶的高效表达

doi:10.3969/j.issn.1673-1689.2016.05.009

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融合短肽促进碱性果胶酶的高效表达
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                        10.3969/j.issn.1673-1689.2016.05.009
                    
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Fusions of Amphipathic Peptide to the Alkaline Polygalacturonate Lyase from Bacillus sp. WSHB04-02 Improves the Production

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摘要:

碱性果胶酶（PGL）是一种重要的工业酶，广泛应用于食品、纺织和造纸等行业。作者将6种双亲短肽（SAP）融合至PGL N端，以期提高PGL在大肠杆菌中的分泌表达效率。得到一株高产菌株PGL-S1，胞外酶活由129.65 U/mL提高到535.47 U/mL，其他5种短肽则酶活降低。通过SDS-PAGE检测发现，PGL-S1的表达量并没有多大变化，推测可能是催化效率提高引起酶活提高。分析双亲短肽的疏水性指数发现，SAP1的疏水性不同于其他短肽，疏水性是蛋白质发生聚集的主要作用力，能够固定N、C末端，加强与底物的相互作用。推测PGL-S1融合蛋白的表面疏水性提高，促进了对底物的催化效率，具有巨大的工业应用潜力。

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Alkaline polygalacturonatelyase（PGL）is one of important industrial enzymes，which was widely used in food，textile and paper industries. In this study，we constructed and expressed six Self-assembling amphipathic peptides （SAPs），whihch were attached to the N terminus of the PGL，to improve the secretory expression of PGL in E.coli. Finally PGL-S1 stood out with enzyme activity improving from 129.65 U/mL to 535.47 U/mL，while the otherenzyme activity weredecreased. There was no obvious change in the extracellular yield of PGL by SDS-PAGE analysis. It indicated that the high enzyme activity was caused by the improving catalytic efficiency. The Grave Value showed that the hydrophobicity of SAP1 was different with the others，which was the main interaction for the accumulation of protein to enhance the interaction between protein and substrate. It was conferred that the improving surface hydrophobicity made a big effect on the catalytic efficiency. PGL-S1has huge potential use for industrial application.

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汪明星,刘松,刘龙,堵国成,陈坚.融合短肽促进碱性果胶酶的高效表达[J].食品与生物技术学报,2016,35(5):504-509.

WANG Mingxing, LIU Song, LIU Long, DU Guocheng, CHEN Jian. Fusions of Amphipathic Peptide to the Alkaline Polygalacturonate Lyase from Bacillus sp. WSHB04-02 Improves the Production[J]. Journal of Food Science and Biotechnology,2016,35(5):504-509.

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在线发布日期: 2016-11-01
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