Mlc A method of high cell-density culture of Escherichia coli by overproducing Mlc protein

摘要

PURPOSE: A method for high cell-density culture of Escherichia coli by over producing Mlc protein is provided, thereby the mutant strain capable of over expressing Mlc protein can be used for increasing the amount of a recombinant protein produced. CONSTITUTION: The method for over-expressing Mlc protein comprises inducing mutation of a 10 region in the transcription-controlling regions and the Shine-Dalgarno(SD) sequence, which is a ribosome-binding site, of the making large colonies(mlc) gene in E. coli, in which the method may further comprise mutation of a transcription-initiating codon of the mlc gene from GTG to ATG; the 10 region is a region from 13 to 8 of the mlc gene sequence and is substituted with a nucleotide sequence of TATAAT; the SD sequence is a sequence from +24 to +31 of the mlc gene sequence and is substituted with a nucleotide sequence of TAAGGAGGA; and the mlc gene sequence has the nucleotide sequence represented by SEQ ID NO: 1. A mutant SR754 (KFCC 11244) contains the mlc gene having mutated transcription-controlling region.