摘要：【中文】本发明公开了一种大肠杆菌诱导蛋白表达的发酵方法，属于生物技术领域。该发酵方法包括以下步骤：将待发酵的种子液接种于发酵培养液中，并置于0.05~0.1MPa的压力条件下进行恒温发酵培养6~10h后，得到发酵底液；往上述发酵底液中添加诱导剂；所述诱导剂在所述发酵底液中质量浓度为0.1~1.5mmol/L；往上述加有诱导剂的发酵底液中添加补料液，并继续进行恒温发酵培养20~48h，得到发酵产物；所述补料液与所述加有诱导剂的发酵底液的体积比为(0.5~2):1。本发明提供的发酵方法可以在36~40℃的环境下进行诱导蛋白的表达，其无需对发酵底液进行快速降温，也可以达到与降温到25~30℃的蛋白表达量，从而可以大大降低能耗以及节约生产成本。 【EN】The invention discloses a fermentation method for expressing escherichia coli induced protein, and belongs to the technical field of biology. The fermentation method comprises the following steps: inoculating a seed solution to be fermented into a fermentation culture solution, and performing constant-temperature fermentation culture for 6-10 h under the pressure condition of 0.05-0.1 MPa to obtain a fermentation base solution; adding an inducer to the fermentation base solution; the mass concentration of the inducer in the fermentation base solution is 0.1-1.5 mmol/L; adding a feed liquid into the fermentation base liquid added with the inducer, and continuously carrying out constant-temperature fermentation culture for 20-48 h to obtain a fermentation product; the volume ratio of the feed liquid to the fermentation base liquid added with the inducer is (0.5-2): 1. The fermentation method provided by the invention can be used for inducing protein expression in an environment of 36-40 ℃, does not need to rapidly cool the fermentation base liquid, and can reach the protein expression amount of cooling to 25-30 ℃, thereby greatly reducing energy consumption and saving production cost.

摘要：【中文】本发明提供一种工业用酶液的热处理澄清方法，包括以下步骤：(1)选择目的蛋白培养发酵：选择目的蛋白，按照现有发酵生产工艺发酵得到发酵液，再将发酵液进行细胞破碎处理，即得细胞破碎液；(2)热处理澄清：将步骤(1)所得细胞破碎液进行热处理澄清；(3)后处理：将步骤(2)热处理澄清后的液体进行离心处理，并收集清液，即为目的蛋白的工业用酶澄清液。该方法具有工艺简单、成本较低且具有环保优势，澄清后酶液中目的蛋白的酶活损失不大于10％，且浊度低于500NTU并能够通过0.2μm的过滤膜。 【EN】The invention provides a heat treatment clarification method of industrial enzyme liquid, which comprises the following steps: (1) selecting target protein for culture fermentation: selecting target protein, fermenting according to the existing fermentation production process to obtain fermentation liquor, and then performing cell disruption treatment on the fermentation liquor to obtain cell disruption liquid; (2) and (3) heat treatment clarification: carrying out heat treatment clarification on the cell disruption solution obtained in the step (1); (3) and (3) post-treatment: and (3) centrifuging the liquid subjected to heat treatment and clarification in the step (2), and collecting clear liquid, namely the industrial enzyme clarified liquid of the target protein. The method has the advantages of simple process, low cost and environmental protection, the enzyme activity loss of the target protein in the clarified enzyme solution is not more than 10 percent, the turbidity is lower than 500NTU, and the clarified enzyme solution can pass through a filtering membrane of 0.2 mu m.

摘要：【中文】本发明提供了可用于在工业相关条件下不对称合成β‑羟基‑α‑氨基酸的工程化醛缩酶多肽。本公开内容还提供了编码工程化醛缩酶多肽的多核苷酸、能够表达工程化醛缩酶多肽的宿主细胞和使用工程化醛缩酶多肽产生β‑羟基‑α‑氨基酸的方法。与其他制备方法相比，使用本发明的工程化醛缩酶多肽用于β‑羟基‑α‑氨基酸的制备，所得产物中立体异构体纯度高，反应条件温和，具有很好的工业应用前景。 【EN】The present disclosure also provides polynucleotides encoding the engineered aldolase polypeptides, host cells capable of expressing the engineered aldolase polypeptides, and methods of producing β -hydroxy- α -amino acids using the engineered aldolase polypeptides.