5 results about "Specific substrate" patented technology

This invention discloses a glycosyltransferase mutant and its application in the synthesis of rebaudioside. Based on the amino acid sequence of the glycosyltransferase shown in SEQ ID NO:1, this invention performs single-point and multi-point mutations to obtain mutants with enhanced catalytic activity. The catalytic activity, substrate specificity, and / or substrate specificity of the glycosyltransferase mutant are altered; mutations at specific sites can significantly improve the enzyme's catalytic activity for specific substrates. The mutant enzyme is obtained by inducing expression and purifying the protein from the obtained mutant sequence. Using the mutant enzyme as a catalyst and UDPG as a glycosyl donor, the catalytic reaction efficiency for the substrates steviol glycoside ST, rebaudioside A (RebA), and rebaudioside D (RebD) can be significantly improved. The glycosyltransferase UGT76G1 mutant constructed in this study improves its catalytic activity, and the efficient production of rebaudioside M is achieved through optimization of the reaction system.

The present application relates to the field of bioengineering, and particularly relates to genome, expression module, recombinant strain and application thereof in preparation of rhodiolin. The present application provides a genome comprising one or more of expression gene, knockout gene, overexpression gene and inhibition gene. The present application constructs a strain capable of utilizing sugarcane molasses as the only carbon source to generate rhodiolin, thereby reducing the cost of materials; then the metabolic network of the microorganism is restructured through metabolic engineering, the metabolic flow is optimized, the metabolic flux among different metabolic modules is reasonably distributed, and the efficient synthesis of the target product is promoted; finally, the mining and screening of enzymes are crucial, the enzymes with higher activity and higher specificity for specific substrates are mined and screened based on the combination of the trained large language model and the machine learning model and the structural information of the substrates, which significantly improves the titer and conversion rate of rhodiolin generation, and at the same time, reduces the generation of by-products and the accumulation of precursor substances, and will promote the large-scale production and application of rhodiolin.

The present application belongs to the technical field of biotechnology and enzyme engineering, and relates to a molecular modification method for improving substrate specificity of aldehyde ketone reductase, which is achieved by constructing a substrate library of aldehyde ketone reductase AKR13B3 and analyzing kinetic parameters, determining a mutation site by performing loop structure analysis and amino acid sequence alignment on a complex structure of aldehyde ketone reductase AKR13B3, coenzyme NADPH and substrate 3-keto-DON, and screening a mutant with increased substrate specificity, the present application designs the geometric shape of the entrance loop region of the substrate binding pocket of AKR13B3 to change substrate preference and realize the increase of catalytic activity of the enzyme on specific substrates, in addition, the present application provides a feasible scheme for designing aldehyde ketone reductase with good substrate specificity, and provides great hope for developing high-efficiency enzyme preparations.

Provided is a method of controlling a substrate transfer system transferring substrates by controlling a first transfer device and a second transfer device comprising: a process of acquiring process module-specific substrate transfer positions for each of the plurality of process modules, wherein the process module-specific substrate transfer positions are substrate transfer positions of the mounting parts of the first transfer device for allowing the second transfer device to place substrates at centers of the stages, the second transfer device transferring substrates from the mounting parts of the load lock module to the stages of the process modules.

The invention relates to improved limonene synthases for biosynthetic production of limonene. engineered limonene synthases with improved characteristics compared to reference limonene synthases are provided. Some engineered limonene synthases have improved limonene production yield. Other engineered limonene synthases have switched enantioselectivity. Still other engineered limonene synthases have improved yield with specific substrates. Some engineered limonene synthases have improvements in a combination of one or more of limonene production yield, enantioselectivity and substrate specificity. Also provided herein is a method of screening limonene synthase, e.g., for characterization of limonene synthases. Further provided is a method of producing limonene using the engineered limonene synthases provided herein.