Method for detecting activity of short-chain isopentenyl transferase

The invention discloses a method for detecting the activity of short-chain isopentenyl transferase. The method comprises the following steps: step 1, carrying out in-vitro expression and purification on short-chain isopentenyl transferase recombinant protein; step 2, establishing a short-chain isope...

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Hauptverfasser:	TANG QIAN, LU SHAN, SONG SHUYAN, LYU DEXIN
Format:	Patent
Sprache:	chi ; eng
Schlagworte:	BEER BIOCHEMISTRY CHEMISTRY COMPOSITIONS OR TEST PAPERS THEREFOR COMPOSITIONS THEREOF CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES CULTURE MEDIA ENZYMOLOGY INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES MEASURING MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING ORGANIC CHEMISTRY PEPTIDES PHYSICS PROCESSES OF PREPARING SUCH COMPOSITIONS PROCESSES USING MICROORGANISMS PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SPIRITS TESTING VINEGAR WINE
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Beschreibung
Zusammenfassung:	The invention discloses a method for detecting the activity of short-chain isopentenyl transferase. The method comprises the following steps: step 1, carrying out in-vitro expression and purification on short-chain isopentenyl transferase recombinant protein; step 2, establishing a short-chain isopentenyl transferase reaction system; step 3, adding alkaline phosphatase to hydrolyze the product of the short-chain isopentenyl transferase; step 4, extracting and concentrating the isopentenyl alcohol compound dissolved in the water phase by using an organic solvent; and 5, carrying out GC-MS analysis on the product to realize qualitative analysis on the to-be-detected substance. All short-chain isopentenyl transferase products can be detected at the same time, purified short-chain isopentenyl transferase recombinant protein can be efficiently obtained in vitro, and qualitative and quantitative analysis of corresponding products is achieved. 本发明公开了一种短链异戊烯基转移酶活性检测方法，包括以下步骤：步骤1，对短链异戊烯基转移酶重组蛋白进行体外表达与纯化；步骤2，建立短链异戊烯基转移