摘要
产油酵母的油脂合成是高度耗氧的生物过程,为提高油脂合成过程中细胞利用氧气的能力,将透明颤菌的血红蛋白基因vgb整合到油脂高产菌株圆红冬孢酵母(Rhodosporidium toruloides,R.toruloides)的染色体中。首先构建了透明颤菌血红蛋白基因vgb表达载体pZPK-PPGK-hyg-TNOS-PGPD-vgb-THSP,再通过农杆菌介导转化的方法将其转化至R.toruloides NP11中,获得重组菌株。Western blot和CO差异光谱分析表明,透明颤菌血红蛋白在R.toruloides NP11中成功表达并且具有生物活性。摇瓶发酵结果表明:与出发菌株相比,在氧气充足条件下,重组菌株NG-21的油脂产量提高了18%;在氧气不足条件下,重组菌株NG-22的油脂产量提高了28%;在限氧和不限氧的条件下,透明颤菌血红蛋白均能够促进圆红冬孢酵母的油脂积累。
Lipid biosynthesis is highly oxygen-demanding in oleaginous yeast,therefore,the VHb expression cassette was integrated into the Rhodosporidium toruloides NP11 chromosome to improve the ability of utilize oxygen via the Agrobacterium-mediated transformation.Firstly,the recombinant expression plasmid pZPK-PPGK-hyg-TNOS-PGPD-vgb-THSP was constructed,and transformed into R.toruloides NP11 by Agrobacterium-mediated transformation to obtain the recombinant strains R.toruloides NG-20,NG-21 and NG-22.The western blot analysis and carbon monoxide(CO)difference spectrum were carried to verify the function of VHb and shown to be biochemically active.The shake flask fermentation showed that the lipid yield of the engineering strain NG-21 was 18%higher than that of the original strain under aerobic condition,the engineering strain NG-22 was 28%higher than that of the original strain under microaerobic condition.In this study,it had been proved that VHb could be able to promote the lipid accumulation in R.toruloides under microaerobic condition.
作者
王爽
张月
周人辉
吕力婷
钱斯日古楞
张素芳
赵宗保
Wang Shuang;Zhang Yue;Zhou Renhui;Lv Liting;Qiansiriguleng;Zhang Sufang;Zhao Zongbao(School of Biological Engineering,Dalian Polytechnic University,Dalian 116034,China;Division of Biotechnology,Dalian Institute of Chemical Physics,CAS,Dalian 116023,China)
出处
《可再生能源》
CAS
北大核心
2020年第9期1143-1148,共6页
Renewable Energy Resources
基金
国家自然科学基金项目(31870042)
中国科学院大连化学物理研究所创新基金(DICP I201947)。
作者简介
通讯作者:张素芳(1978-),女,博士,副研究员,研究生导师,研究方向为产油酵母分子微生物学和代谢工程。E-mail:zsfjxy@dicp.ac.cn。
