Preparation method of noradrenaline based on L-threonine aldolase R318L/H128N

Preparation method of noradrenaline based on L-threonine aldolase R318L/H128N

According to the method, a mutant R318L/H128N of L-threonine aldolase derived from a metagenome database of black bear intestinal microorganisms is coupled with tyrosine decarboxylase derived from enterococcus faecium, glycine and 3, 4-dihydroxybenzaldehyde are used as substrates, and the nor-adrena...

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Hauptverfasser: QI NA, ZHA RENFEN, WANG BOCHU, LEI BIJING, MA JINGSONG
Format: Patent
Sprache:chi ; eng
Schlagworte:
BEER
BIOCHEMISTRY
CHEMISTRY
COMPOSITIONS THEREOF
CULTURE MEDIA
ENZYMOLOGY
FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE
METALLURGY
MICROBIOLOGY
MICROORGANISMS OR ENZYMES
MUTATION OR GENETIC ENGINEERING
PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS
SPIRITS
VINEGAR
WINE
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Zusammenfassung:According to the method, a mutant R318L/H128N of L-threonine aldolase derived from a metagenome database of black bear intestinal microorganisms is coupled with tyrosine decarboxylase derived from enterococcus faecium, glycine and 3, 4-dihydroxybenzaldehyde are used as substrates, and the nor-adrenaline is synthesized through double-enzyme coupling. The enantioselectivity of the noradrenaline is 97%, and the yield of the noradrenaline reaches 76%. The amino acid sequence of the L-threonine aldolase mutant R318L/H128N is as shown in SEQ ID NO: 3, and the L-threonine aldolase mutant R318L/H128N is obtained by changing the 318th amino acid of L-threonine aldolase of which the amino acid sequence is SEQ ID NO: 1 into Leu from Arg and changing the 128 amino acid of L-threonine aldolase of which the amino acid sequence is SEQ ID NO: 1 into Asn from His; the amino acid sequence of the tyrosine decarboxylase is as shown in SEQ ID NO: 5. The double-enzyme coupling method for synthesizing noradrenaline has the advantag