The invention provides a highly active S-cyanohydrin lyase obtained by mutating an amino acid residue at position 103 of a wild-type cassava S-cyanohydrin lyase. The mutation can significantly increase an expression of a mutant enzyme in E. coli and does not require a decrease in temperature when induced. Further mutations at position 128 and other sites were performed to obtain mutants with increased catalytic activity.

A method for preparation of and use of R-hydroxynitrile lyase derived from mango. Specifically, a variety of mutant cyanohydrin lyases were obtained by constructing random mutation and site-saturation mutagenesis libraries and high-throughput screening, and the method was safe, simple and easy to operate.

The present disclosure relates to a multi-enzyme conjugate, a method for preparing the same and a method for preparing an organic compound using the same. More particularly, a multi-enzyme conjugate exhibiting improved catalytic efficiency over respective free enzymes using site-specific incorporation of a clickable non-natural amino acid into the enzymes and two compatible click reactions, a method for preparing the same and a method for preparing an organic compound using the same may be provided.

The invention provides a highly active S-cyanohydrin lyase obtained by mutating an amino acid residue at position 103 of a wild-type cassava S-cyanohydrin lyase. The mutation can significantly increase an expression of a mutant enzyme in E. coli and does not require a decrease in temperature when induced. Further mutations at position 128 and other sites were performed to obtain mutants with increased catalytic activity.

Provided are a method for obtaining an HNL gene and HNL derived from a millipede other than Chamberlinius hualienensis, and preparing a practically useable amount of HNL; and a method for producing optically active cyanohydrin using this HNL. A method for producing a millipede-derived HNL gene. A method that includes the selection of a gene having a base sequence that encodes a conserved amino acid sequence TAX1DIX2G (SEQ ID NO: 15) or VPNGDKIH (SEQ ID NO: 16) of millipede-derived HNL from genes present in an organism belonging to the Diplopoda. A protein having an amino acid sequence of any of (1)-(3) and having HNL activity. (1) An amino acid sequence listed in any of SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13, 83, 85, 87, or 89; (2) an amino acid sequence having amino acids deleted, substituted, and/or added in an amino acid sequence of (1); or (3) an amino acid sequence having 90% or greater identity to an amino acid sequence of (1). A method for preparing optically active cyanohydrin by causing this millipede-derived HNL to act on a reaction solvent that contains an aldehyde or the like and a substance that generates hydrogen cyanide or the like.

The present invention is directed to methods of modifying the plant development process comprising of exposing a plant or plant part to volatiles biosynthesized by one or more bacteria or enzymes. Specifically, the embodiment uses one or more bacteria selected from the plant growth promoting bacteria group consisting of Rhodococcus spp., Pseudomonas spp., Bacillus spp., or Xanthobacter spp., or a mixture thereof. A closed apparatus, FIG. 1A, containing a tri-phasic system is used to expose the bacteria to hydrocarbons, iron, cyanide, and/or ammonium compounds; the method induces the biocatalyst to biosynthesize volatile compound(s) that deter ethylene production in climacteric plants or fruit resulting in the biocatalyst ability to delay fruit ripening.

The present disclosure relates to a multi-enzyme conjugate, a method for preparing the same and a method for preparing an organic compound using the same. More particularly, a multi-enzyme conjugate exhibiting improved catalytic efficiency over respective free enzymes using site-specific incorporation of a clickable non-natural amino acid into the enzymes and two compatible click reactions, a method for preparing the same and a method for preparing an organic compound using the same may be provided.

Methods for synthesizing and using R-hydroxynitrile lyase derived from mango. Specifically, a variety of mutant cyanohydrin lyases were obtained by mutating a wild-type hydroxynitrile lyase gene, adding an enzyme cleavage site thereto, preparing a recombinant plasmid by inserting the mutated wild-type hydroxynitrile lyase gene into a vector, introducing the recombinant plasmid into a strain of bacteria, and secreting and expressing the R-hydroxynitrile lyase in the strain. The method Is safe, simple and easy to operate.

The present invention is directed to methods of modifying the plant development process comprising of exposing a seed, seedling, or plant to volatiles biosynthesized by one or more bacteria or enzymes. Specifically, the embodiment uses one or more bacteria selected from the plant growth promoting bacteria group consisting of Rhodococcus spp., Pseudomonas spp., Bacillus spp., or Xanthobacter spp., or a mixture thereof. A closed apparatus, FIG. 1A, containing a tri-phasic system is used to expose the bacteria to hydrocarbons, iron, cyanide, and/or ammonium compounds; the method induces the biocatalyst to biosynthesize volatile compound(s) that plant hormone production to enhance or accelerate plant development.

Provided are a method for obtaining an HNL gene and HNL derived from a millipede other than Chamberlinius hualienensis, and preparing a practically useable amount of HNL; and a method for producing optically active cyanohydrin using this HNL. A method for producing a millipede-derived HNL gene. A method that includes the selection of a gene having a base sequence that encodes a conserved amino acid sequence TAX1DIX2G (SEQ ID NO: 15) or VPNGDKIH (SEQ ID NO: 16) of millipede-derived HNL from genes present in an organism belonging to the Diplopoda. A protein having an amino acid sequence of any of (1)-(3) and having HNL activity. (1) An amino acid sequence listed in any of SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13, 83, 85, 87, or 89; (2) an amino acid sequence having amino acids deleted, substituted, and/or added in an amino acid sequence of (1); or (3) an amino acid sequence having 90% or greater identity to an amino acid sequence of (1). A method for preparing optically active cyanohydrin by causing this millipede-derived HNL to act on a reaction solvent that contains an aldehyde or the like and a substance that generates hydrogen cyanide or the like.