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黄曾慰,梁达奉,曾练强,吴兆鹏,马步,常国炜.朱黄青霉α-葡聚糖酶在毕赤酵母中的高效表达[J].广西科学,2014,21(6):614-618. [点击复制]
- HUANG Zeng-wei,LIANG Da-feng,ZENG Lian-qiang,WU Zhao-peng,MA Bu,Chang Guo-wei.High Expression of Dextranase from Penicillium Minioluteum in Pichia pastoris[J].Guangxi Sciences,2014,21(6):614-618. [点击复制]
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| 朱黄青霉α-葡聚糖酶在毕赤酵母中的高效表达 |
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黄曾慰1, 梁达奉1,2, 曾练强1, 吴兆鹏1, 马步2, 常国炜1
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| (1.广州甘蔗糖业研究所, 广东省甘蔗改良与生物炼制重点实验室, 广东广州 510316;2.广西农垦糖业集团股份有限公司, 广西糖业研发中心, 广西南宁 530002) |
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| 摘要: |
| [目的]提高朱黄青霉(Penicillium minioluteum)α-葡聚糖酶(Dextranase)基因在毕赤酵母(Pichia pastoris)中的表达水平。[方法]根据毕赤酵母的密码子偏爱对酶基因进行优化与合成。优化后的基因片段与载体pGAPZαA连接,转化毕赤酵母KM71H。[结果]获得组成型分泌表达α-葡聚糖酶的工程菌KM71H/pGAPZαA-dex。发酵工艺试验中,摇瓶培养144 h,酶活为153 U/mL。6.8 L发酵罐补料分批培养92 h,酶活达到1218 U/mL。[结论]该工程菌以甘油作为碳源,发酵调控简单,产酶水平较高,具有适用于大规模生产的潜力。 |
| 关键词: α-葡聚糖酶 密码子优化 毕赤酵母 重组菌 |
| DOI： |
| 投稿时间：2014-10-10修订日期：2014-11-13 |
| 基金项目:现代农业产业技术体系建设专项项目(CARS-20-4-5)和八桂学者建设工程专项经费项目资助。 |
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| High Expression of Dextranase from Penicillium Minioluteum in Pichia pastoris |
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HUANG Zeng-wei1, LIANG Da-feng1,2, ZENG Lian-qiang1, WU Zhao-peng1, MA Bu2, Chang Guo-wei1
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| (1.Guangzhou Sugarcane Industry Research Institute, Guangdong Key Lab of Sugarcane Improvement & Biorefinery, Guangzhou, Guangdong, 510316, China;2.Guangxi State Farms Sugar Industrial Group Company Limited, Guangxi Sugarcane Industry R & D Center, Nanning, Guangxi, 530002, China) |
| Abstract: |
| [Objective] The aim of this paper is to enhance the expression level of a dextranase gene dex of Penicillium minioluteum in Pichia pastoris.[Methods] Codon optimization was applied and the dextranase gene was synthesized using the preferential condon of pichia pastoris.The codon-optimized gene was cloned into the vector pGAPZαA and transformed into host strain KM71H to achieve constitutive expression and secretion of dextranase.[Results] The activity of dextranase reached 153 U/mL after 144 h when the engineering strain KM71H/pGAPZαA-dex was cultured in shake flask.Using glycerol as the sole carbon source, a simple fermentation control strategy has been developed and expression level of 1218 U/mL after 92 h has been demonstrated in 6.8 L scale fed-batch fermentation.[Conclusion] The high expression level makes the engineering strain a good candidate for industrial production. |
| Key words: dextranase codon optimization Pichia pastoris recombinant strain |
