Abstract: To provide an improved ?-fructofuranosidase which is capable of efficiently producing kestose while inhibiting the production of nystose. This improved ?-fructofuranosidase comprises either: an amino acid sequence (a) obtained by introducing, into the amino acid sequence represented by SEQ ID NO: 2, an amino acid mutation i) in which the 85th glycine (G) from the N-terminal is substituted for a protein-constituting amino acid other than glycine (G), and/or an amino acid mutation ii) in which the 310th histidine (H) from the N-terminal is substituted for lysine (K), arginine (R), or tyrosine (Y); or an amino acid sequence (b) which exhibits ?-fructofuranosidase activity, and which comprises an amino acid sequence obtained by deleting, substituting, inserting, or adding one or more amino acids in (a) other than the amino acid into which the amino acid mutation has been introduced.

Abstract: An improved ?-fructofuranosidase is provided. The improved ?-fructofuranosidase may comprise an amino acid sequence having 60% or higher identity to the amino acid sequence of SEQ ID NO: 2, and may contain an amino acid mutation that replaces histidine (H) corresponding to position 395 counted from the N terminus of SEQ ID NO: 2 with arginine (R) or lysine (K), an amino acid mutation that replaces leucine (L) at position 123 counted from the N terminus of SEQ ID NO: 2 with cysteine (C), and/or an amino acid mutation that replaces phenylalanine (F) at position 473 counted from the N terminus of SEQ ID NO: 2 with tyrosine (Y).

Abstract: To provide an improved ?-fructofuranosidase which is capable of efficiently producing kestose while inhibiting the production of nystose. This improved ?-fructofuranosidase comprises either: an amino acid sequence (a) obtained by introducing, into the amino acid sequence represented by SEQ ID NO: 2, an amino acid mutation i) in which the 85th glycine (G) from the N-terminal is substituted for a protein-constituting amino acid other than glycine (G), and/or an amino acid mutation ii) in which the 310th histidine (H) from the N-terminal is substituted for lysine (K), arginine (R), or tyrosine (Y); or an amino acid sequence (b) which exhibits ?-fructofuranosidase activity, and which comprises an amino acid sequence obtained by deleting, substituting, inserting, or adding one or more amino acids in (a) other than the amino acid into which the amino acid mutation has been introduced.

Abstract: [Problem] To provide a production method capable of simply and efficiently producing a fructose-added carbohydrate using ?-fructofuranosidase. [Solution] A method for producing a fructose-added carbohydrate, said method having a step in which a receptor substrate and a hydrate containing terminal fructose residue are brought into contact with: Escherichia coli expressing an anchor protein for expression on a cell surface and ?-fructofuranosidase as one polypeptide, a composition including dead cells of the expressing Escherichia coli, or a polypeptide obtained from the expressing Escherichia coli and including an amino acid sequence of ?-fructofuranosidase.

Abstract: A method for producing cis-5-hydroxy-L-pipecolic acid is described. A gene recombinant microorganism enabling direct production of cis-5-hydroxy-L-pipecolic acid can be used in the method. Also described is a gene recombinant microorganism. In particular, it is described that a gene recombinant microorganism having DNAs encoding proteins involved in the biosynthesis of L-pipecolic acid and a DNA encoding a protein having the L-pipecolic acid cis-5-hydroxylase activity is cultured in a medium, and cis-5-hydroxy-L-pipecolic acid is collected from the medium.

Abstract: [Problem] To provide an improved ?-fructofuranosidase which enables the production of kestose with high efficiency while reducing the amount of a by-product such as nystose produced during the production of kestose. [Solution] An improved ?-fructofuranosidase comprising an amino acid sequence which is produced by introducing an amino acid mutation into an amino acid sequence for a ?-fructofuranosidase having 60% or higher identity to the amino acid sequence for wild-type ?-fructofuranosidase which is represented by SEQ ID NO: 2, wherein the amino acid mutation is such a mutation that, when amino acid sequence alignment is performed, a histidine (H) residue corresponding to the position 395 as numbered from the N-terminal of the amino acid sequence for the wild-type ? fructofuranosidase which is represented by SEQ ID NO: 2 can be replaced by an arginine (R) residue or a lysine (K) residue.

Abstract: [Problem] To provide a production method capable of simply and efficiently producing a fructose-added carbohydrate using ?-fructofuranosidase. [Solution] A method for producing a fructose-added carbohydrate, said method having a step in which a receptor substrate and a hydrate containing terminal fructose residue are brought into contact with: Escherichia coli expressing an anchor protein for expression on a cell surface and ?-fructofuranosidase as one polypeptide, a composition including dead cells of the expressing Escherichia coli, or a polypeptide obtained from the expressing Escherichia coli and including an amino acid sequence of ?-fructofuranosidase.

Abstract: A method for producing cis-5-hydroxy-L-pipecolic acid is described. A gene recombinant microorganism enabling direct production of cis-5-hydroxy-L-pipecolic acid can be used in the method. Also described is a gene recombinant microorganism. In particular, it is described that a gene recombinant microorganism having DNAs encoding proteins involved in the biosynthesis of L-pipecolic acid and a DNA encoding a protein having the L-pipecolic acid cis-5-hydroxylase activity is cultured in a medium, and cis-5-hydroxy-L-pipecolic acid is collected from the medium.

Abstract: An expression vector capable of expressing a foreign gene in Pseudonocardia autotrophica; a transformant of Pseudonocardia autotrophica produced by using the expression vector; a method for producing a protein by using the transformant; a method for producing an active form of vitamin D3 from vitamin D3, which comprises highly expressing a gene encoding an enzyme involved in the synthesis of the active form of vitamin D3 in a transformant by using the expression vector or the transformant; a method for producing 25-hydroxyvitamin D2 from vitamin D2; and a method for producing pravastatin from compactin, which comprises highly expressing a compactin hydroxylase gene in a transformant by using the expression vector or the transformant.

Abstract: Disclosed is a means for improving the poor conversion efficiency in a conventional bioconversion system using a transformant which is given by introducing a gene originated from xerogenic organisms. A transformant is prepared by using a host which is defective in a gene encoding a multidrug efflux protein and introducing a gene originated from xerogenic organisms. Use of the transformant results in much effective microbial conversion of a hydrophobic or amphipathic substrate compound into a desired compound. In case, an Escherichia coli is used as the host, the gene encoding a multidrug efflux protein to be defective may be tolC, acrA, acrB and the like.

Abstract: Disclosed is a means for improving the poor conversion efficiency in a bioconversion system using an Escherichia coli cell having a bacterium-originated cytochrome P-450 gene integrated therein. A recombinant Escherichia coli cell is produced by introducing aciB and aciC which encode a gene for the electron transport system originated from the Acinetobacter sp. OC4 strain into an Escherichia coli cell, and adding a polynucleotide encoding an N-terminal sequence composed of 48 amino acid residues of AciA and the like to the 5?-terminus of a bacterium-originated cytochrome P-450 gene, wherein AciA is an alkane-oxidative cytochrome P-450 originated from the Acinetobacter sp. OC4 strain. Use of the recombinant Escherichia coli cell results in much effective microbial conversion of a hydrophobic or amphipathic substrate compound into a desired compound.

Abstract: The invention offers a method of hydrogenating camptothecin in inert solvent in the presence of nickel catalyst, whereby selective hydrogenation of camptothecin can be very efficiently accomplished using the low-cost catalyst.

Abstract: A vitamin D3 hydroxylase is purified from Pseudonocardia autotrophica cell, and a primer is designed based on amino acid sequence obtained from hydroxylase. Subsequently, PCR is conducted using genomic DNA of Pseudonocardia autotrophica as a template to clone a gene for the vitamin D3 hydroxylase. By conducting a conversion reaction using a microorganism in which the vitamin D3 hydroxylase gene is expressed using a proper expression system, a hydroxide of vitamin D or the like (e.g., hydroxy vitamin D3) can be produced with high efficiency.