• Microbiology and Biotechnology Letters
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• v.18 no.5
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• pp.530-534
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• 1990

A strain of actinomycetes, Thermoactinomyces sp. CH-53, was isolated from manure and composted livestock feces. Actinornycetes-feed additive was prepared with the solid wheat bran medium of Thermoactinomyces sp. CH-53 that grew vigorously on unsterilized poultry feces at $50^{\circ}C$. pH 6.5- 9.5 and moisture content of 55-65% and added at a rate of 1% (wtlwt) to the commercially assorted feed to be fed poultry. The excreted feces contained $10^7-10^8$. Thermoactinomyces sp. CH-53 cells per gram. Poultry feces malodour was got rid of during treatment. The effect on plant growth was evaluated on the basis of the amount of nitrogen as fertilizer under a loading of 0.2g N1600g soillpot. A11 samples were showed a promotion effect for plant growth. The treated poultry feces added from O.lg to 0.4g total nitrogen per 600g soil in a pat increased the growth of Brassica rapa var. perui-ridis.

• Microbiology and Biotechnology Letters
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• v.28 no.3
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• pp.167-171
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• 2000

Analysis of Production of Thermostable Alkaline Protease using Thermoactinomyces sp. E79. Jung, Sang Won, Sung-Sik Park, Yong-Cheol Park" Tae Kwang Oh2, and Jin-Ho Seo*, Department of Food Science and Technology, Seoul National University, Suwon 441-744, Korea, 1lnterdisciplinary program [or Biochemical Engineering & Biotechnology, Seoul National Univer5it}~ Seoul 151 "7421 Koreal 2Microbial Enzyme RU, Korea Research Institute of Bioscience & Biotechnology, Po. Box 1151 Yusong, Taejon 305"6001 Korea - This research was undertaken to analyze fermentation properties of Thermoactinomyces sp. E79 for production of a thermostable alkaline protease, which is able to specifically hydrolyze defatted soybean meal (DSM) to amino acids. TIle optimum pH for cell growth and protease production was pH 6.7, Thermoactinomyces sp. E79 did not grow at pHlO Among carbon sources tested, soluble starch was the best for protease production, while glucose repressed protease production. Tryptone was found to be the best nitrogen source for cell growth and soytone was good tor protease production. Oxygen transfer rate played an important role in producing thermostable alkaline protease. Ma'<..imum values of 6.58 glL of dry cell weight and 43.0 UJmL of protease activity were obtained in a batch fermentation using a 2.5 L jar fermentor at 1.93 X 102 hr-l of volumetric oxygen transfer coeff'jcient (kLa). Addition of 200 mgIL humic acid to the growth medium resulted in 1.64 times higher protease activity and 1.77 times higher cell growth than the case without humic acid addition.

• Journal of Microbiology and Biotechnology
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• v.27 no.12
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• pp.2190-2198
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• 2017

Thermoactinomyces sp. strain YT06 was isolated from poultry compost and observed to degrade integral chicken feathers completely at $60^{\circ}C$, resulting in the formation of 3.24 mg/ml of free amino acids from 50 ml of culture containing 10 g/l chicken feathers. Strain YT06 could grow and secrete keratinase using feather as the only carbon and nitrogen sources without other supplement, but complementation of 10 g/l sucrose and 4 g/l $NaNO_3$ increased the production of the keratinolytic enzyme. The maximum protease activity obtained was 110 U/ml and for keratinase was 42 U/ml. The keratinase maintained active status over a broad pH (pH 8-11) and temperature ($60-75^{\circ}C$). It was inhibited by serine protease inhibitors and most metal ions; however, it could be stimulated by $Mn^{2+}$ and the surfactant Tween-20. A reductive agent (${\beta}$-mercaptoethanol) was observed to cleave the disulfide bond of keratin and improve the access of the enzyme to the keratinaceous substrate. Zymogram analysis showed that strain YT06 primarily secreted keratinase with a molecular mass of approximately 35 kDa. The active band was assessed by MALDI-TOF mass spectrometry and was observed to be completely identical to an alkaline serine protease from Thermoactinomyces sp. Gus2-1. Thermoactinomyces sp. strain YT06 shows great potential as a novel candidate in enzymatic processing of hard-to-degrade proteins into high-value products, such as keratinous wastes.

• Journal of Microbiology and Biotechnology
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• v.9 no.4
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• pp.469-474
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• 1999

Thermoactinomyces sp. E79 produced two types of thermostable alkaline proteases extracellularly. A minor protease was separated from a major protease by using DEAE-column chromatography. This enzyme was purified to homogeneity by ammonium sulfate and DEAE-Sepharose ion-exchange chromatography. The purified minor protease showed different biochemical properties compared to the major protease. The molecular mass of the purified enzyme was estimated by SDS-PAGE to be 36 kDa. Its optimum temperature and pH for proteolytic activity against Hammarsten casein were $70^{\circ}C$ and 9.0, respectively. The enzyme was stable up to$75^{\circ}C$ and in an alkaline pH range of 9.0-11.0. The enzyme was inhibited by phenylmethylsulfonyl fluoride (PMSF) and $Hg^{2+}, indicating that the enzyme may be a cysteine-dependent serine protease. In addition, the enzyme cleaved the endoproteinase substrate, succinyl-Ala-Ala-Pro-Phe-p- nitroanilide, and the $K_m$ value for the substrate was 1.2 mM.

• Proceedings of the Microbiological Society of Korea Conference
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• 1998.04a
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• pp.54.2-54.2
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• 1998

• Journal of Microbiology and Biotechnology
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• v.8 no.6
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• pp.560-567
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• 1998

We have found that Thermoactinomyces vulgaris KFB-Cl00 produces a chitinase. The optimum temperature and pH of the enzyme activity were $55^{\circ}C$ and 6.5. The enzyme was stable after heat treatment at $80^{\circ}C$ for 30 min and stable in acidic and basic conditions (PH 6.0~11.0). The thermostable endo-chitinase from Thermoactinomyces vulgaris KFB-C100 was cloned into the plasmid pBR322 by using E. coli DH5$\alpha$ as a host strain. The positive clone carrying a recombinant plasmid (PKCHI23) with a 4.1-kb fragment containing the chitinase gene was found. The recombinant plasmid was analyzed to determine the essential region for chitinase activity and obtained a 2.3-kb fragment, which was sub cloned into pTrc99A using the PstI and SalI sites to construct pTrc99A/pKCHI23-3. The resulting plasmid exerted high chitinase activity upon transformation of E. coli XL1-Blue cells. Chitinase was overproduced 14 times more in the clone cells than in the wild-type cells and the enzyme was purified to homogeneity. The purified enzyme showed the similar properties as the native chitinase from T. vulgaris in terms of molecular weight and substrate specificity. The catalytic action of the cloned enzyme was an endo type, producing chitobiose as a major reaction product.

• Journal of Microbiology and Biotechnology
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• v.11 no.1
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• pp.85-91
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• 2001

To construct an efficient Bacillus subtilis expression vector, strong promoters were isolated from the chromosomal DNA libraries of Clostridium acetobutylicum ATCC 4259, Thermoactinomyces sp. E79, and Bacillus thermoglucosidasius KCTC 3400. The $P_{C27}$ promoter cloned from the clostridial chromosmal DNA showed a 5-fold higher promoter strength than the $P_{SP02}$ promoter in the expression of the cat gene, and its sequence was estimated as an upstream region of the predicted hypothetical gene (tet-R family bacterial transcription regulator gene) in C. acetobutylicum. As a promoter element, $P_{C27}$ exhibited putative nucleotide sequences that can bind with bacterial RNAP and the 3'end of the 16S rRNA just upstream of the start codon. In addition, the promoter activity of $P_{C27}$ was distinctively repressed in the presence of glucose. Using $P_{C27}$ as the promoter element, a glucose controllable B. subtilis expression vector was constructed and the lipase gene from Staphylococcus haemolyticus KCTC 8957P was expressed in B. subtilis. When compared with the lipase expression by the T7 promoter induced by IPTG in E. coli, the $P_{C27}$ promoter showed about a 1.5-fold higher expression level in B. subtilis than that without induction.

• The Microorganisms and Industry
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• v.27 no.1
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• pp.2-17
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• 2001

Cyclomaltodextrinase(CDase, EC 3.2.1.54), maltogenic amylase(EC 3.2.1.133). neopullulanase(EC 3.2.1.135)는 cyclomaltodextrin(CD), pullulan 및 전분을 가수분해하는 효소들이다. 이 효소들은 $\alpha$-1,4-Ο-glycosidic 결합에 작용하여 CD와 전분을 말토오스로 pullulan을 panose로 가수분해할 뿐만 아니라 올리고당들을 다양한 당 수용체 분자들의 C-3, C-4. C-6 수산기로 전이시키는 활성도 갖고 있다. 이러한 특성들은 기존의 $\alpha$-amylase를 비롯한 판수화물 분해효소들과 뚜렷이 구별되는 것으로 전분 분해효소들의 분류체계에 새로운 기준점을 제시한다고 하겠다. 본 총설에서는 CDase, maltogenic amylase, neopullulanase처럼 pullulan이나 전분보다 CD를 훨씬 더 잘 분해하는 효소들과 Thermoactinomyces vulgaris amylase II(TVA II)처럼 CD를 분해하기는 하나 pullulan을 더 잘 분해하는 효소들의 생화학적, 효소적, 구조적 특성들을 종합하여 소개하고자 하였다. 이 효소들은 40~60% 정도로 아미노산 서열이 동일하고, 세포 내에 존재하며, 분자량이 62~90 kDa로 $\alpha$-amylase보다 다소 크다. 아미노산 서열 비교분석 및 maltogenic amylase와 TVA II 등의 3차구조 분석 결과, 이 효소들은 아미노 말단에 보통 $\alpha$-amylase에는 존재하지 않는 약 130개 아미노산으로된 영역을 갖고 있어 이를 매개로 이합체를 형성할 수 있는 것으로 나타났다. 이합체-단위체 평형은 염 농도, 효소 농도, 산도 등에 의해 조절되고 단위체와 이합체 모두 효소환성을 갖고 있으나, 기질 특이성이 다르며 단위체는 전분을, 이합체는 CD를 선호하는데 이는 이합체 형성 시 활성부위의 구조적 변화에 따른 것으로 분석되었다. 본 총설에서는 CD 분해효소들의 다양한 기질 특이성을 올리고머 형성 등의 구조적 특성과 관련하여 논함으로써 관련 효소들의 분류체계를 보다 명확히 할 수 있는 자료를 제공하고자 하였으며, 이러한 효소들의 생리적 기능 및 산업적 이용에 대해 제안하고자 하였다.

• Korean Journal of Soil Science and Fertilizer
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• v.29 no.3
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• pp.303-311
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• 1996

The microflora changes of 10 water-controled treatments combined with livestock manures(pig, chicken) and bulking agents(sawdust, paper sludge) were investigated. The B/F values of the P-1 and C-1(65%, $H_2O$) treatments were 3571 and 5400 respectively, but those of the P-4 and C-4(50%, $H_2O$) treatments showed very low values, 667 and 334, respectively. The B/F values tended to increase with higher water content of the treatments. In the composting processes, the successions of microflora, adapting the compost environments, took place via fluctuating temperature. In the high temperature period, the numbers of mesophilic bacteria and fungi decreased, but that of the spore forming bacteria increased. However, the number of mesophilic bacteria inereased during the cold period. The B/F values of compost ranged 25-300, which indicates a decrease in the quantity of bacteria. The time required for the temperature of compost to reach $60^{\circ}C$ showed different patterns. There was no pathogenic microorganism in the treatments which reached a high temperature in a short period of time, but, in the treatments which reached a high temperature over a Long period of time, the pathgenic microorganism was not still alive.