Mussel adhesive proteins (MAPs) have great potential as bioglues, particularly in wet conditions. Although in vivo residue‐specific incorporation of 3,4‐dihydroxyphenylalanine (Dopa) in tyrosine‐auxotrophic Escherichia coli cells allows for production of Dopa‐incorporated bioengineered MAPs (d MAPs), the low production yield hinders the practical application of d MAPs. This low production yield of d MAPs is due to low translational activity of a noncanonical amino acid, Dopa, in E. coli cells. Herein, to enhance the production yield of d MAPs, we investigated the coexpression of Dopa‐recognizing tyrosyl‐tRNA synthetases (TyrRSs). To use the Dopa‐specific Methanococcus jannaschii TyrRS (MjTyrRS‐Dopa), we altered the anticodon of tyrosyl‐tRNA amber suppressor into AUA (MjtRNA^Tyr_AUA) to recognize a tyrosine codon (AUA). Co‐overexpression of MjTyrRS‐Dopa and MjtRNA^Tyr_AUA increased the production yield of Dopa‐incorporated MAP foot protein type 3 (d fp‐3) by 57%. Similarly, overexpression of E. coli TyrRS (EcTyrRS) led to a 72% higher production yield of d fp‐3. Even with coexpression of Dopa‐recognizing TyrRSs, d fp‐3 has a high Dopa incorporation yield (over 90%) compared to ones prepared without TyrRS coexpression.

中文翻译：

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使用工程翻译机器增强掺入多巴的贻贝粘附蛋白的生产。

贻贝粘附蛋白 (MAP) 作为生物胶具有巨大的潜力，尤其是在潮湿条件下。尽管酪氨酸营养缺陷型大肠杆菌细胞中 3,4-二羟基苯丙氨酸 (Dopa) 的体内残基特异性掺入允许生产掺入多巴的生物工程 MAPs ( d MAPs)，但产量低阻碍了d MAPs的实际应用。d MAPs 的这种低产量是由于非经典氨基酸多巴在大肠杆菌细胞中的低翻译活性。在此，为了提高d MAPs 的产量，我们研究了多巴识别酪氨酰-tRNA 合成酶（TyrRSs）的共表达。使用多巴特定的对于詹氏甲烷球菌TyrRS (MjTyrRS-Dopa)，我们将酪氨酰-tRNA 琥珀抑制基因的反密码子更改为 AUA (MjtRNA ^Tyr _AUA ) 以识别酪氨酸密码子 (AUA)。MjTyrRS-Dopa 和 MjtRNA ^Tyr _{AUA 的}共同过表达使掺入多巴的 MAP 足蛋白 3 型 ( d fp-3) 的产量提高了57%。同样，大肠杆菌TyrRS (EcTyrRS) 的过表达导致d fp-3 的产量提高了 72% 。即使与多巴识别 TyrRSs 共表达，与没有 TyrRS 共表达的情况相比，d fp-3 也具有高多巴掺入率（超过 90%）。