Kanokkorn Sinma

Shizuoka University, Sizuoka, Shizuoka, Japan

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Publications (6)2.01 Total impact

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    ABSTRACT: An extracellular thermostable xylanase produced by Saccharopolyspora pathumthaniensis S582 was purified 167-fold to homogeneity with a recovery yield of 12%. The purified xylanase appeared as a single protein band on SDS-PAGE, with a molecular mass of 36 kDa. The optimal temperature and pH of the xylanase were 70 °C and 6.5. The enzyme was stable within a pH range of 5.5-10.0. It retained its activity after incubation at 50 °C for 2 h. Its half lives at temperatures of 60 and 70 °C were 180 and 120 min respectively. Hydrolysis of beechwood xylan by the xylanase yielded xylobiose and xylose as major products. The enzyme acted specifically on xylan as an endo-type xylanase, and exhibited a K(m) value of 3.92 mg/mL and a V(max) value of 256 µmol/min/mg. Enzyme activity was completely inhibited by Hg(2+), and was stimulated by Rb(+) and Cs(+). The xylanase gene was cloned from genomic DNA of Saccharopolyspora pathumthaniensis S582 and sequenced. The ORF consisted of 1,107 bp and encoded 368 amino acid residues containing a putative signal peptide of 23 residues. This xylanase is a new member of family (GH) 10 that shows highest identity, of 63.4%, with a putative xylanase from Nocardiopsis dassonvillei subsp. dassonvillei.
    No preview · Article · Oct 2011 · Bioscience Biotechnology and Biochemistry
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    ABSTRACT: Morphological and chemotaxonomic characterization of actinomycete strain S582 isolated from the gut of a termite (Speculitermes sp.) in Pathum Thani Province, Thailand, clearly demonstrated that this strain is a member of the genus Saccharopolyspora. 16S rDNA sequence analysis for the strain supported the assignment of the strain to the genus Saccharopolyspora. The similarity value of sequences between this strain and the closely related species Saccharopolyspora endophytica was 99.5%. The DNA G+C content was 70.2 mol%. DNA-DNA hybridization results (53.3%) and some physiological and biochemical properties indicated that strain S582(T) was distinguished from the phylogenetically closest relatives. Based on these genotypic and phenotypic data, strain S582(T) should be a new species in the genus Saccharopolyspora and the name Saccharopolyspora pathumthaniensis sp. nov. is proposed for the strain. The type strain is S582(T) (=NBRC 104112(T) =BCC 28624(T)).
    Full-text · Article · Apr 2011 · The Journal of General and Applied Microbiology
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    ABSTRACT: Actinomycetes are a group of prokaryotic organisms belonging to Gram-positive bacteria and play an important ecological role in recycling substances in the nature. To determine possible established correlation between isolated actinomycetes and its biochemical degradation (xylan, soluble starch, cassava, protein, lipid, uric acid, carboxymethyl cellulose and avicel degradation) and oxidation property (guaiacol), the actinomycetes were isolated from Termes sp. The dendogram was generated from UPGMA analysis with FreeTree software. In this work, 45 strains of actinomycetes were isolated from guts of Termes sp. Among these, 44 isolated strains could degrade protein, soluble starch and cassava starch. Twenty-three strains degraded uric acid and xylan. The isolated strains that able to degrade avicel and oxidize lignin were rare. The morphological character showed the variety of aerial hyphae and spore forming in each strain. The dendogram was constructed based on biodegradation activity of tested strains. The tested strains were classified into 5 clusters. Cluster 1, cluster 2, cluster 3, cluster 4 and cluster 5 contained 14, 9, 9, 9 and 4 isolated strains, respectively. The actinomycetes strains showed a similar biodegration activity within a same cluster. This result indicated the relation between biodegradation activity and actinomycetes strains. Dendogram based on the biodegradation activity was found to be an efficiencient tool for grouping purpose.
    No preview · Article · Apr 2011
  • K Khucharoenphaisan · K Sinma
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    ABSTRACT: The strain PNR11 was isolated from gut of termite during the screening for uric acid degrading actinomyces. This strain was able to produce an intracellular uricase when cultured in fermentation medium containing uric acid as nitrogen source. Base on its morphological characters and 16S rDNA sequence analysis, this strain belong to the genus Saccharopolyspora. This is the first report ofuricase produced from the genus Saccharopolyspora. The aim of this study was to investigate the effects of different factors on uricase production by new source of Saccharopolyspora. Saccharopolyspora sp. PNR11 was cultured in production medium in order to determine the best cultivation period. The result showed that the time period required for maximum enzyme production was 24 h on a rotary shaker operating at 180 rpm. Optimized composition of the production medium consisted of 1% yeast extract, 1% maltose, 0.1% K2HPO4, 0.05% MgSO4 7H2O, 0.05% NaCl and 1% uric acid. The optimum pH and temperature for uricase production in the optimized medium were pH 7.0 and 30 degrees C, respectively. When the strain was cultured at optimized condition, the uricase activity reached to 216 mU mL(-1) in confidential level of 95%. The crude enzyme had an optimum temperature of uricase was 37 degrees C and it was stable up to 30 degrees C at pH 8.5. The optimum pH ofuricase was 8.5 and was stable in range of pH 7.0-10.0 at 4 degrees C. This strain might be considered as a candidate source for uricase production in the further studies. Present finding could be fulfill the information ofuricase produce from actinomycetes.
    No preview · Article · Feb 2011 · Pakistan Journal of Biological Sciences
  • K. Khucharoenphaisan · K. Sinma
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    ABSTRACT: Xylanase is useful enzyme in various industrial such as conversion of lignocellulose to fermentable sugars for the production of chemicals and biofuels. Thermomyces lanuginosus is a potent thermophilic strain which produces high level of cellulose-free xylanase. The aim of this study was to investigate the effects of signal sequence on T. lanuginosus xylanase expression in Escherichia coli. The xylanase gene from those was amplified with and without signal sequence and expressed in E. coli. The result showed that the transformant (pUC/Xynsig) containing original signal sequence from T. lanuginosus SKR produced xylanase with 56% higher than that of another transformant (pUC/Xyn) lacking of original signal sequence. The recombinant enzymes were partially purified using ammonium sulfate precipitation and DEAE cellulose column and then determined some properties. The optimal pH and optimal temperature of xylanase from pUC/Xynsig was pH 6.0 and 50°C, respectively. In case of pUC/Xyn without signal sequence, the values of xylanase showed higher than those having pH 7.0 and temperature of 70°C in which was similar to original host. In conclusion, original signal sequence from T. lanuginosus SKR could increase xylanase production when expressed in E. coli but some properties of expressed enzyme were changed. However, this finding could apply to other expression system of various hosts in order to stimulate the level of the protein production.
    No preview · Article · Feb 2011 · Biotechnology(Faisalabad)
  • K Khucharoenphaisan · K Sinma
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    ABSTRACT: Thermomyces lanuginosus is thermophilic fungus in which was isolated from widespread material. A high number of this fungus was found in composts especially mushroom composts. This fungus has been reported to produce a high level xylanase when cultivated in the medium containing xylan and corn cob as a carbon source. Various strains of T. lanuginosus produced a single xylanase with molecular masses in range of 22.0 to 29.0 kDa. Pure beta-xylanase obtained from various strains of this fungus exhibited highly stability at high temperature and wide pH range. The optimal temperature and optimal pH of pure beta-xylanase from various strains of T. lanuginosus have been reported in range of 60-75 degrees C and pH 6.0-7.0, respectively. The great thermal stability was resulting from the present of hydrophilic amino acid on beta sheet of the surface of xylanase structure. Moreover, the relatedness between high and low xylanase producing strains can be distinguish by random amplification of polymorphic DNA (RAPD). Based on nucleotide sequences and T. lanuginosus xylanase gene has been classified to be a member of family 11 (formerly known as cellulase family G) glycosyl hydrolases. This enzyme was endo-type xylanase having main product are xylose and xylobiose. The expression of xylanase gene from T. lanuginosus was achieved in Escherichia coli and methylotrophic yeast Pichia pastoris. The ability of T. lanuginosus in which produced large amount of high thermos stable xylanase has made this fungus to be a source of xylanase production for biobleaching in pulp and paper process.
    No preview · Article · Jun 2010 · Pakistan Journal of Biological Sciences