Escherichia colt L - asparaginase was modified with O - carboxymethylated chitosan using glutaraldehyde as a coupling agent. The resulting coujugate retained more than 50% of its original enzyme activity under theprot...Escherichia colt L - asparaginase was modified with O - carboxymethylated chitosan using glutaraldehyde as a coupling agent. The resulting coujugate retained more than 50% of its original enzyme activity under theprotection of its normal substrate or product and shoWed marked resistance toproteolysis by trypsin and chymotrypsin.展开更多
Recombinant E. coli BL 21 was cultivated in high cell density to produce human-like collagen. The effects of the feeding of nitrogen source, controlled by an auto on/off-feeding mode with two different cycles of 0.5mi...Recombinant E. coli BL 21 was cultivated in high cell density to produce human-like collagen. The effects of the feeding of nitrogen source, controlled by an auto on/off-feeding mode with two different cycles of 0.5min and 4min intervals, oxygen-enrichment methods and inducement strength on the cell yield and human-like collagen production were investigated. The studies showed that nitrogen source feeding in fast cycle could result in higher human-like collagen production than that in slow cycle; and the feedback regulation of glucose, increase of the pressure of fermentation bioreactor, and supply of oxygen-enriched air could all increase cell yield and human-like collagen production. The effects of inducement strength on protein expression were found important. When OD600 reached 90-100, the cultivation temperature was increased to 42℃ to begin induction for 2-3 h, and then shifted to 39℃ for 5-6h induction, the cell density and human-like collagen production could reach 96g·L-1 [DCW (dry cell mass)] and 19.8% (g·g-1 DCW) respectively.展开更多
文摘Escherichia colt L - asparaginase was modified with O - carboxymethylated chitosan using glutaraldehyde as a coupling agent. The resulting coujugate retained more than 50% of its original enzyme activity under theprotection of its normal substrate or product and shoWed marked resistance toproteolysis by trypsin and chymotrypsin.
基金Supported by the National Science and Technology Key Funds (2003DA901A32)the National Nature Science Foundation (No. 20476085).
文摘Recombinant E. coli BL 21 was cultivated in high cell density to produce human-like collagen. The effects of the feeding of nitrogen source, controlled by an auto on/off-feeding mode with two different cycles of 0.5min and 4min intervals, oxygen-enrichment methods and inducement strength on the cell yield and human-like collagen production were investigated. The studies showed that nitrogen source feeding in fast cycle could result in higher human-like collagen production than that in slow cycle; and the feedback regulation of glucose, increase of the pressure of fermentation bioreactor, and supply of oxygen-enriched air could all increase cell yield and human-like collagen production. The effects of inducement strength on protein expression were found important. When OD600 reached 90-100, the cultivation temperature was increased to 42℃ to begin induction for 2-3 h, and then shifted to 39℃ for 5-6h induction, the cell density and human-like collagen production could reach 96g·L-1 [DCW (dry cell mass)] and 19.8% (g·g-1 DCW) respectively.
