Alkaline feed

摘要

A method for cultivating a bacterial cell comprising the addition of an amino acid in an alkaline solution used for pH regulation. Also an aspect is a method for producing a polypeptide comprising the steps of a) providing a bacterial cell comprising a nucleic acid encoding the polypeptide, b) cultivating the provided cell, c) adjusting the pH value during the cultivating with a basic solution comprising an amino acid, d) recovering the polypeptide from the cell or the cultivation medium and thereby producing the polypeptide.

主权项

1. A method for cultivating an Escherichia coli cell expressing a polypeptide wherein the cultivating comprises adding an alkaline solution of an amino acid selected from aspartate, aspartic acid, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, tryptophan and tyrosine during the cultivating;
wherein the amino acid has a concentration in the alkaline solution that is higher than its solubility in water at 20° C. and at neutral pH, and the amino acid has a concentration of 30 g/l or more; wherein the alkaline solution is an ammonia solution of 10% (w/v) or more; and wherein the dry cell weight of the cultivated bacterial cell is at one point in the cultivating at least 20 g/l; wherein the polypeptide is an apolipoprotein A1 polypeptide having an amino acid sequence selected from SEQ ID NO: 01 through 35.