Publication:
Characterization of poly(L-lactide)-degrading enzyme produced by thermophilic filamentous bacteria Laceyella sacchari LP175

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Issued Date

2014

Resource Type

Article

File Type

application/pdf

ISSN

221260

DOI

10.2323/jgam.60.13

Other identifier(s)

2-s2.0-84896473825

Rights Holder(s)

Scopus

Bibliographic Citation

Journal of General and Applied Microbiology. Vol 60, No.1 (2014), p.13-22

Suggested Citation

Hanphakphoom S., Maneewong N., Sukkhum S., Tokuyama S., Kitpreechavanich V. Characterization of poly(L-lactide)-degrading enzyme produced by thermophilic filamentous bacteria Laceyella sacchari LP175. Journal of General and Applied Microbiology. Vol 60, No.1 (2014), p.13-22. doi:10.2323/jgam.60.13 Retrieved from: https://hdl.handle.net/20.500.14740/6442

Title

Characterization of poly(L-lactide)-degrading enzyme produced by thermophilic filamentous bacteria Laceyella sacchari LP175

Author(s)

Hanphakphoom S.
 Maneewong N.
 Sukkhum S.
 Tokuyama S.
 Kitpreechavanich V.

Abstract

Eleven strains of poly(L-lactide) (PLLA)-degrading thermophilic bacteria were isolated from forest soils and selected based on clear zone formation on an emulsified PLLA agar plate at 50°C. Among the isolates, strain LP175 showed the highest PLLA-degrading ability. It was closely related to Laceyella sacchari, with 99.9% similarity based on the 16S rRNA gene sequence. The PLLA-degrading enzyme produced by the strain was purified to homogeneity by 48.1% yield and specific activity of 328 U·mg-protein-1 with a 15.3-fold purity increase. The purified enzyme was strongly active against specific substrates such as casein and gelatin and weakly active against Suc-(Ala)<inf>3</inf>-pNA. Optimum enzyme activity was exhibited at a temperature of 60°C with thermal stability up to 50°C and a pH of 9.0 with pH stability in a range of 8.5-10.5. Molecular weight of the enzyme was approximately 28.0 kDa, as determined by gel filtration and SDS-PAGE. The inhibitors phenylmethylsulfonyl fluoride (PMSF), ethylenediaminetetraacetate (EDTA), and ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA) strongly inhibited enzyme activity, but the activity was not inhibited by 1 mM 1,10-phenanthroline (1,10-phen). The N-terminal amino acid sequences had 100% homology with thermostable serine protease (thermitase) from Thermoactinomyces vulgaris. The results obtained suggest that the PLLA-degrading enzyme produced by L. sacchari strain LP175 is serine protease. © 2014 Applied Microbiology, Molecular and Cellular Biosciences Research Foundation.

Subject(s)

1,10 phenanthroline
 Benzylsulfonyl fluoride
 Casein
 Edetic acid
 Egtazic acid
 Gelatin
 RNA 16S
 Serine proteinase
 Thermitase
 Bacterial protein
 Bacterial RNA
 Enzyme
 Polyester
 Polylactide
 RNA 16S
 Amino acid sequence
 Amino terminal sequence
 Article
 Bacterial strain
 Bacterium isolate
 Bacterium isolation
 Controlled study
 DNA sequence
 Enzyme activity
 Enzyme analysis
 Enzyme inhibition
 Enzyme purification
 Enzyme specificity
 Enzyme synthesis
 Gel filtration
 Gene sequence
 Laceyella sacchari
 Molecular weight
 Nonhuman
 Nucleotide sequence
 PH measurement
 Polyacrylamide gel electrophoresis
 Sequence homology
 Temperature sensitivity
 Thermoactinomyces
 Thermoactinomyces vulgaris
 Thermophilic bacterium
 Thermostability
 Bacillales
 Bacterial gene
 Bioremediation
 Chemistry
 Enzyme stability
 Enzymology
 Genetics
 Heat
 Isolation and purification
 Metabolism
 Microbiology
 Molecular genetics
 Phenotype
 Phylogeny
 RNA gene
 Scanning electron microscopy
 Amino Acid Sequence
 Bacillales
 Bacterial Proteins
 Base Sequence
 Biodegradation, Environmental
 Enzyme Stability
 Enzymes
 Genes, Bacterial
 Genes, rRNA
 Hot Temperature
 Microscopy, Electron, Scanning
 Molecular Sequence Data
 Phenotype
 Phylogeny
 Polyesters
 RNA, Bacterial
 RNA, Ribosomal, 16S
 Soil Microbiology
 Substrate Specificity

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URI

https://hdl.handle.net/20.500.14740/6442

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Scopus: Year 1983-2021

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