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rDNA介导的菊粉酶基因整合载体构建及在K.marxianus中应用

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Indexed by:期刊论文

Date of Publication:2013-03-15

Journal:大连理工大学学报

Included Journals:PKU、ISTIC、CSCD、Scopus

Volume:53

Issue:2

Page Number:176-182

ISSN No.:1000-8608

Key Words:菊粉酶基因;整合表达;Kluyveromyces marxianus;菊芋;乙醇

Abstract:为提高K luyverom yces marxianus(K.marxianus)发酵菊芋粉生产乙醇过程中的菊粉酶活性,以酿酒酵母rDNA为整合位点,构建菊粉酶基因的整合表达载体pFA6a-rDNA-pgk-inu.经SphⅠ线性化后,采用电击法转化K.marxianus,使表达载体整合到K.marxianus的染色体上.经PCR鉴定的阳性转化子能够分泌菊粉酶且能在无筛选压力下稳定遗传50代.重组菌K/r-2分泌的菊粉酶活力为140 U/mL,比出发菌株提高了80%.以粗菊芋粉生料为底物进行了补料发酵,重组菌株的发酵终点乙醇浓度为76.5 g/L,比出发菌株提高了5 g/L,达到理论转化率的96%.这些研究工作为非粮作物菊芋生产燃料乙醇奠定了基础.

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