US5139935A
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Method of regulating expression of a foreign gene by controlling the sugar concentration in a medium and a process of producing a foreign product thereby
Abstract Escherichia coli carrying a hybrid plasmid having been constructed by inserting a desired foreign gene into an expression vector so as to permitting expression of said desired foreign gene therein was cultured in a medium containing a sugar component utilizable by the E. coli as a carbon source at a concentration of 0.3% or more so that the expression of said desired foreign gene was suppressed. This E. coli (i.e., transformant) was cultured in the medium in which the sugar concentration was maintained at 0.3% or more in a first process so as to supress the suppression of the foreign gene and to support sufficient cell growth and thereafter at less than 0.3% in a second process to release the suppression of the expression so as to permit effective production of the foreign gene product, which resulted in high concentration of the foreign gene product in the final culture.
A process for producing L-phenylalanine ammonialyase derived from Rhodosporidium toruloides by culturing an Escherichia coli carrying a promoter which is either a P L lambda promoter or a combined promoter containing both the tac promoter and the P L promoter, and a foreign structural gene encoding for L-phenylalanine ammonialyase derived from Rhodosporidium toruloides linked to said promoter so as to permit expression of said foreign structural gene under the direction of said promoter, which process comprises steps of: (a) culturing said E. coli in a culture medium containing a sugar component and a carbon source, in which the concentration of the sugar component is maintained at least 0.3%, and thereafter (b) culturing said E. coli grown in said first culture process, in which the concentration of said sugar component in the second culture medium is maintained at less than 0.3%.
Claim 2
Dependent Claim
140 words
A process for producing L-phenylalanine ammonialyase derived from Rhodosporidium toruloides by culturing an Escherichia coli carrying a promoter which is either a P L lambda promoter or a combined promoter containing both the tac promoter and the P L promoter, and a foreign structural gene encoding for L-phenylalanine ammonialyase derived from Rhodosporidium toruloides linked to said promoter so as to permit expression of said foreign structural gene under the direction of said promoter, which process comprises steps of: (a) culturing said E. coli in a culture medium containing a sugar component and a carbon source, in which the concentration of the sugar component is maintained at least 0.3%, and thereafter (b) culturing said E. coli grown in said first culture process, in which the concentration of said sugar component in the second culture medium is maintained at less than 0.3%.
Claim 3
Dependent Claim
26 words
A process of producing a foreign gene product as set forth in claim 1, wherein said L-phenylalanine ammonialyase has the amino acid sequence as follows: ##STR1##
Claim 4
Dependent Claim
35 words
A process of producing a foreign gene product as set forth in claim 1, wherein said E. coli is E. coli MT-10424 which is obtained by transforming Escherichia coli MC1061 with plasmid pSYP L -3.
Claim 5
Dependent Claim
33 words
A process of producing a foreign gene product as set forth in claim 1, wherein said E. coli is E. coli MT-10423 which is obtained by transforming Escherichia coli MC1061 with plasmid pSW115.