来自Trichoderma longibrachiatum UN32的氨基转移酶TlBCAT的表征,它参与了树突蛋白类型总化物生物合成

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Xu Qian¹, Tuifan Yu¹, Yiwen Cao²

¹College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing, Jiangsu, 211800, PR China.

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|November 8, 2024

概括

分支链氨基酸氨基转移酶 (BCATs) 在Trichoderma longibrachiatum UN32.中对登德罗宾类总类 (DTTAs) 的产生至关重要. 补充外源链氨基酸可以增强真菌生长和DTTA产量.

科学领域:

生物化学生物化学
分子生物学分子生物学
菌群学菌群学菌群学是指菌群学

背景情况:

特里科德尔玛长腕 (Trichoderma longibrachiatum UN32) 在工业上产生相关的登德罗宾类型总化物 (DTTA).
分支链氨基酸氨基转移酶 (BCATs) 参与二次代谢产物.
化治疗之前在T. longibrachiatum UN32.32中增强了DTTAs和TlBCAT基因表达.

研究的目的:

从T. longibrachiatum UN32.32中克隆和表征TlBCAT基因.
研究TLBCAT的酶活性和基质特异性.
探索外源链氨基酸 (BCAA) 在T. longibrachiatum UN32生长和DTTAs生产中的作用.

主要方法:

基因克隆和TlBCAT的测序.
重组TLBCAT的蛋白质表达和净化.
酶分析测试以确定催化效率和基质特异性.
分析BCAA对真菌生长和DTTAs生产的影响.

主要成果:

TlBCAT的cDNA为1191 bp,编码了一个397氨基酸蛋白 (约. 42 kDa) 的时间.
重组TLBCAT有效催化L-氨酸转胺,其催化效率为15.91mM-1S-1.1.
TlBCAT对分支链和疏水性氨基酸具有广泛的基质特异性,利用α-甲酸作为氨基接受剂.
在PLP结合口袋中的高度保存的残留物表明其具有功能意义.
外源BCAA显著促进了T. longibrachiatum UN32的生长和DTTAs的产生.

结论:

TlBCAT是T. longibrachiatum UN32.32中DTTAs生物合成中的一个关键酶.
酶的广泛基质特异性对于其在二次新陈代谢中的作用至关重要.
用外源BCAA补充提供了一种可行的策略,以提高真菌培养中的DTTA产量.

关键词:

三皮长 (Trichoderma longibrachiatumatum) 是一种植物.分支链氨基酸氨基转移酶是什么丹德罗宾类型的总化物.引起引起的引起的

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