• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.509.1-509
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• 1986

Recent advances in recombinant DNA techniques have provided a tool for breeding of microorganisms of hyper production. Enzyme production by cloned microorganism has some advantages. They are ⅰ) Enzymes can be produced by a microorganism easily cultured ⅱ) Hyper production. ⅲ) In some cases, such as thermophilic enzyme gene is cloned in a mesophilic bacteria, the enzyme purification procedure can be simplified. One example, production of thermophilic ${\beta}$-galactosidase in B. subtilis will be presented. Bacillus stearothermophilus IAM 11001 produced three ${\beta}$-galactosidases, ${\beta}$-galactosidase I, II and III (${\beta}$-gal-I, II and III). By connecting restriction fragments of the chromosomal DNA to plasmid vector, followed by transformation of Escherichia coli, two ${\beta}$-galactosidase genes (bgaA and bgaB) located close to each other on the chromosome were cloned.

• Proceedings of the Korean Society of Life Science Conference
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• 2002.03a
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• pp.83-83
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• 2002

• Journal of Microbiology and Biotechnology
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• v.10 no.2
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• pp.244-250
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• 2000

An ${\alpha}-glucosidase$ gene (aglA) from thermophilic Bacillus sp. DG0303 was cloned, sequenced, and expressed in Escherichia coli. The aglA was localized to the 2.1-kb PvuI-XmnI region within the 5.9-kb DNA insert of the gybrid plasmid pAG1. The gene consisted of an open reading frame of 1,686 bp with an unusual GTG initiation codon and TGA termination codon. The amino acid sequence deduced from the nucleotide sequence predicted a protein of 562 amino acid residues with a M, of 66,551 dalton. A comparative amino acid sequence analysis revealed that DG0303 ${\alpha}-glucosidase$ is related to bacillary oligo-1, 6-glucosidases. The Bacillus sp. DG0303 ${\alpha}-glucosidase$ showed a high sequence identity (36-59%) to the B. flavocaldarius, B. cereus, and B. thermoglucosidasius oligo-1, 6-glucosidases. The number of prolines in theses four ${\alpha}-glucosidases. was observed to increase with increasing thermostability of these enzymes. The cloned ${\alpha}-glucosidase was purified from E. coli $DH5{\alpha}$ bearing pAG1 and characterized. The recombinant enzyme was identical with the native enzyme in its optimum pH and in its molecular mass, estimated by sodium dodecy1 sulfate-polyacrylamide gel electrophoresis. The temperature optimum of the cloned ${\alpha}-glucosidase$ was lower than that of the native enzyme.

• Journal of Life Science
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• v.9 no.5
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• pp.510-517
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• 1999

For the treatment of Korean food-wastes, three mesophilic and one thermophilic bacteria were isolated from soil and fermented fertilizers. The thermophilic Streptomyces sp. strain WF021 produced two enzymes which were a protease and a lipase at 55$^{\circ}C$. The mesophilic Bacillus sp. strain WF024 produced four enzymes which were a protease, a lipase, a amylase and a cellulase when the strain was grown both at 3$0^{\circ}C$ and 55$^{\circ}C$. The Bacillus sp. PY123 had produced three enzymes which were a protease, a cellulase and a lipase at 3$0^{\circ}C$. The Bacillus sp. strain CM1 produced three enzymes which were a protease, a amylase, and a cellulase at 3$0^{\circ}C$. The bacteria were grown in media containing 6% NaCl at least and did not have antagonism each other. The four isolates treated much more food-wastes than referance strains did. In a flask without aeration, three reference strains treated 15.4% of food-wastes, while four isolates treated 23.7% of food-wastes. In a flask with aeration, food-wastes were treated 67.3% by four isolates, and 64.3% by three reference strains, but 53.9% without bacteria. However, food-wastes were treated about 78% in a 200$\ell$-reactor made by Siwon Co., while 65.8% in a 20$\ell$-reactor made by Sanyo Co.

• KSBB Journal
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• v.9 no.4
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• pp.400-405
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• 1994

The conditions for ${\beta}$-galactosidase production from alkalophilic, thermophilic Bacillus sp. TA-11 were investigated. The maximal enzyme production was obtained when the strain was cultured at $50^{\circ}C$ for 5 days with fed-batch culture in the optimal medium containing 1.5% lactose, 0.6% yeast extract 0.15% $K_2HP0_4$and initial pH 9.5, and then final enzyme activity under the above conditions was 5200 unit/ml of cell free extract.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.5
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• pp.674-680
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• 2015

This study was conducted to characterize the rapidly processed salt-fermented sandfish sauce added Bacillus coagurance KM-1 (RSSS) and commercial salt-fermented sandfish sauce (CSSS 1, 2). Contents of total nitrogen and amino nitrogen were higher in CSSS 1,2 than in RSSS (P<0.05). Total free amino acid contents of RSSS and CSSS 1,2 were 1,121.2±100 mg/100 g, 1,553.6±98.2 mg/100 g and 1,507.3±99.8 mg/100 g. Major free amino acid of RSSS was glutamic acid (194.4±17.3 mg/100 g), alanine (140.8±12.6 mg/100 g), lysine (135.1±12.1 mg/100 g), leucine (109.8±9.8 mg/100 g), aspartic acid (103.0±9.2 mg/100 g), valine (73.5±6.6 mg/100 g) in ordor. The samples were caused by their composition of the free amino acids rations, in which were umami, sweet and bitter taste in the salt-fermented sandfish sauce during fermentation. The Na was the largest mineral followed by K, Mg, P, Ca in the samples (P<0.05). Sensory evaluation result of samples, CSSS 1 was the highest than the others in overall acceptance.

• Animal Bioscience
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• v.36 no.4
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• pp.671-678
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• 2023

Objective: Previous studies isolated the thermophilic ammonium-tolerant (TAT) bacterium Bacillus sp. TAT105 that grew in composting swine manure with the assimilation of ammonium nitrogen and reduced ammonia emissions during composting. Those studies also investigated the potential for applications of TAT105 to composting. It was observed that the concentration of TAT bacteria, phylogenetically close to TAT105, increased during composting. The objectives of this study were to identify the phylogenetic placement of these TAT bacteria and investigate their distribution in various composts. Methods: The phylogenetic placement of TAT105 was examined based on the sequence of 16S ribosomal RNA gene. The genomic DNA homology between TAT105 and the type strains of bacterial species that were phylogenetically close to TAT105 were examined by DNA-DNA hybridization. Moreover, the tolerances of these strains to NH₄Cl and NaCl were analyzed using a cultivation method. Concentrations of TAT bacteria in various composts were evaluated using an agar medium specific to TAT bacteria and polymerase chain reaction followed by restriction fragment length polymorphism analysis. Results: TAT105 was most closely related to Bacillus thermolactis and Bacillus kokeshiiformis. Many variants of these species have been detected in various environments, including composts. The type strains of these species displayed TAT characteristics that were similar to those of TAT105. Among the composts examined in this study, TAT bacteria were detected at high concentrations (10⁵ to 10⁹ colony forming units per gram of dry matter) in most of the composts made from cattle manure, swine manure, bark, and excess sludge. Conclusion: TAT bacteria comprised B. thermolactis, B. kokeshiiformis, and their phylogenetically close relatives. They were considered to be adaptable to composting of some certain materials, and a favorable target for searching for strains with some useful function that could be applied to composting of these materials.

• Environmental Engineering Research
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• v.23 no.3
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• pp.258-264
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• 2018

The objectives of this research were to 1) isolate and identify thermophilic bacteria for food waste treatment; 2) investigate the capability of food waste treatment using Bacillus species; and 3) develop air-dried Bacillus starters for food waste treatment. Five Bacillus species were isolated from food wastes and identified as Bacillus licheniformis (B. licheniformis) G1, Bacillus circulans C2, Bacillus subtilis (B. subtilis) E1, Bacillus vanillea F1, and Bacillus atrophaeus G2 based on 16S rDNA sequencing. Each identified Bacillus and the mixture of Bacillus species were cultivated in the standard food waste at $45^{\circ}C$ for 8 d. Changes in cell count, solid contents, and pH of the food waste were monitored during cultivation. Air-dried Bacillus cell powders were prepared using wheat flour and lactomil as excipients, and the cell count and survival rate were determined. The cell count of B. licheniformis G1 exhibited the highest number among the tested Bacillus (${\sim}10^8CFU/mL$). The greatest reduction in solid contents of food waste was achieved by B. subtilis E1 (22.6%). The mixture of B. licheniformis G1 and B. subtilis E1 exhibited a synergistic effect on the reduction of solid contents. Lactomil was determined as better excipient than wheat flour based on the greatest survival rate of 95%.

• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.363-367
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• 2006

Food wastes were decomposed into the Mugri (Isung Engineering, Korea), a food waste reduction machine, with adding sawdust of cryptomeria. Degradation effects were better when the machine worked at over 45$^{\circ}C$ than those at the lower temperature. Thermophilic bacteria were isolated from cryptomeria sawdust and the food waste products degraded by the machine. The isolates from cryptomeria sawdust were classified into 3 genera (Acinetobacter baumannii, Enterobacter sp. and Erwinia cypripedii) and almost all the isolates from the degraded products were partially identified as Bacillus sp. by 16S rDNA sequence analysis. The isolated thermophilic bacteria showed degradative enzyme activities. In the case of addition of the 30 thermophilic bacteria into the machine, degradation rate of food wastes was almost twice as high with increasing process temperature up to 6$^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.14 no.6
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• pp.447-453
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• 1986

The culture filtrate of thermophilic alkalophilic Bacillus K17 strain contained two types of xylanases were purified by ammonium sulfate fractionation, DEAD-Sephadex A-50 column chromatography, CM-Sephadex C-50 column chromatography and Sephadex G-100 gel filtration. The purified enzymes were found to be homogeneous by sodium dodecyl sulfate and disc polyacrylamide gel electrophoresis. Xylanase I and II were characterized with respect to molecular weight, optimal temperature and pH, thermal and pH stability, and Michaelis constant. Xylanase II was more active and stable, and showed greater substrate affinity and molecular weight than xylanase I. The activities of xylanases I and II were inhibited by Cu$^{++}$, Ag$^+$, Hg$^{++}$ and Fe$^{++}$. Xylanase I hydrolyzed xylan to yield xylobiose and higher amount of xylooligosaccharides, but xylanase II produced xylose other than xylobiose and xylooligosacchrides.