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Engineering the cbh1 Promoter of Trichoderma reesei for Enhanced Protein Production by Replacing the Binding Sites of a Transcription Repressor ACE1 to Those of the Activators.
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2020-01-21 , DOI: 10.1021/acs.jafc.9b05452 Xianhua Sun 1 , Xuhuan Zhang 2 , Huoqing Huang 1 , Yuan Wang 1 , Tao Tu 1 , Yingguo Bai 1 , Yaru Wang 1 , Jie Zhang 1 , Huiying Luo 1 , Bin Yao 1 , Xiaoyun Su 1
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2020-01-21 , DOI: 10.1021/acs.jafc.9b05452 Xianhua Sun 1 , Xuhuan Zhang 2 , Huoqing Huang 1 , Yuan Wang 1 , Tao Tu 1 , Yingguo Bai 1 , Yaru Wang 1 , Jie Zhang 1 , Huiying Luo 1 , Bin Yao 1 , Xiaoyun Su 1
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The strong and inducible cbh1 promoter is most widely used to express heterologous proteins, useful in food and feed industries, in Trichoderma reesei. Enhancing its ability to direct transcription provides a general strategy to improve protein production in T. reesei. The cbh1 promoter was engineered by replacing eight binding sites of the transcription repressor ACE1 to those of the activators ACE2, Hap2/3/5, and Xyr1. While changing ACE1 to Hap2/3/5-binding sites completely abolished the transcription ability, replacements with ACE2- and Xyr1-binding sites (designated cbh1pA and cbh1pX promoters, respectively) largely improved the promoter transcription efficiency, as reflected by expression of a reporter gene DsRed. The cbh1pA and cbh1pX promoters were applied to improve secretory expression of a codon-optimized mannanase from Aspergillus niger to 3.6- and 5.0-fold higher, respectively, which has high application potential in feed industry.
中文翻译：
通过将转录阻遏物ACE1的结合位点替换为激活物的结合位点，对里氏木霉的cbh1启动子进行工程改造，以增强蛋白质的生产。
强大且可诱导的cbh1启动子在里氏木霉中最广泛地用于表达异源蛋白，可用于食品和饲料工业。增强其直接转录的能力提供了改善里氏木霉蛋白质生产的一般策略。通过将转录阻抑物ACE1的八个结合位点替换为激活物ACE2，Hap2 / 3/5和Xyr1的八个结合位点来改造cbh1启动子。虽然将ACE1更改为Hap2 / 3/5结合位点完全取消了转录能力，但用ACE2-和Xyr1结合位点（分别指定为cbh1pA和cbh1pX启动子）进行替换，极大地提高了启动子的转录效率，这反映在报告基因的表达上基因DsRed。
更新日期：2020-01-22
中文翻译：
通过将转录阻遏物ACE1的结合位点替换为激活物的结合位点，对里氏木霉的cbh1启动子进行工程改造，以增强蛋白质的生产。
强大且可诱导的cbh1启动子在里氏木霉中最广泛地用于表达异源蛋白，可用于食品和饲料工业。增强其直接转录的能力提供了改善里氏木霉蛋白质生产的一般策略。通过将转录阻抑物ACE1的八个结合位点替换为激活物ACE2，Hap2 / 3/5和Xyr1的八个结合位点来改造cbh1启动子。虽然将ACE1更改为Hap2 / 3/5结合位点完全取消了转录能力，但用ACE2-和Xyr1结合位点（分别指定为cbh1pA和cbh1pX启动子）进行替换，极大地提高了启动子的转录效率，这反映在报告基因的表达上基因DsRed。
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