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漆酶Lcc9在灰盖鬼伞菌中的同源过表达

ZHANG Jingna, ZHOU Gang,LIU Juanjuan,FANG Zemin,XIAO Yazhong

Journal of Biology(2022)

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Abstract
为了高效制备漆酶Lcc9,克隆了Lcc9编码基因,利用酿酒酵母同源重组技术构建lcc9过表达载体并在Corprinopsis cinerea Okayama 7#130(OK7)和FA2222中进行同源过表达.经原生质体共转化、再生培养基初筛、最简培养基复筛,分别获得46株OK7和29株FA2222阳性转化子.摇瓶发酵结果显示,在Kjalke培养基中接种5%(体积分数),经37℃培养6~7d后,OK7阳性菌株漆酶活力达到5.1~21.1 U/mL,FA2222阳性菌株漆酶活力达到3.7~12.3 U/mL,分别为野生型表达量的19.2和241.2倍.在3 L发酵罐中对FA2222阳性菌株进行漆酶的发酵制备,结果显示,经37℃、50r/min培养108h后菌株漆酶活力达59U/mL,相比普通摇瓶发酵,漆酶活力进一步提高了4.8倍.
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