• 한국미생물·생명공학회지
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• 제28권5호
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• pp.264-269
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• 2000

This study is that of providing a fairly practical practival guide to the use of natural pigment in food industry. A strain isolated from marine resources was carried out the production of red pigment. The pigment showed UV absorption maxima at 520 and 550 nm. The color intensity in aqueous solution was fairly stable in the ranges of pH 5~8. The strain KS-97 produced a maximum yield of red pigment at$ 25^{\circ}C$ for 72 hrs with pH 7.0. The strain KS-97 was iden-tified a Bacillus sp. based on morphological and biochemical characterization such as a rod from, motility, spore for-mation, Gram positive and catalase production. The production of red pigment indicated that the strain Ks-97 utilized at thigh concentration of colloidal chitin as carbon sources obtained maximum yield of red pigment at $25^{\circ}C$ for 72 hrs. The highest production of red pigment was observed with cultivation in medium containing 20% colloi-dal chitin, 2.5g polypeptone, 2.5g yeast extract, 1.0g $KH_2$$PO_4$, 0.01g $MgSO_4$.$6H_2$O, 0.01g $ZnSO_4$, 0.01 g $MnSO_4$(per 1).

• 한국미생물·생명공학회지
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• 제23권2호
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• pp.187-196
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• 1995

In order to produce fuctional chitooligosaccharides, a strain excreting mainly endo-type chitinase suitable for chitooligosaccharides production was newly screened and identified as Aspergillus fumigatus JC-19. The chitinase excretion was repressed in nutrient rich medium but stimulated by colloidal chitin indicating that the chitinase is inducible type enzyme. Maximum secretion of the enzyme was observed at pH 7.0 and 37$\circ$C . The growth and chitinase production patterns of Aspergillus fumigatus JC-19 showed that the cell growth reached maximum after 4-5 days with final chitinase concentration of 0.46 unit per ml. Excreted chitinase was purified by ammonium sulfate precipitation, colloidal chitin adsorption, anion exchange chromatography, and gel filtration, respectively, and measured M.W of 50 KDa. The enzyme reaction carried out both by crude and purified chitinase showed that the purified chitinase accumulated more chitooligosaccharides of 1-6 degree of polymerization than that of crude chitinase.

• 한국미생물·생명공학회지
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• 제26권6호
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• pp.515-522
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• 1998

길항미생물로 분리.동정된 Pseudomonas sp. 3098이 생산하는 chitinase를 황산암모늄 침전, DEAE-cellulose column chromatography, Bio-Gel P-100에 의한 겔여과, 1차 및 2차 hydroxyapatite column chromatography 과정을 거쳐 회수율 5.8%,정제도 15.7배의 정제효소를 얻었고, 효소의 순도는 SDS-PAGE로 확인하였으며, 분자량은 45kDa으로 추정되었다. 정제된 chitinase의 최적 온도와 pH는 45$^{\circ}C$와 5.0이었고, 정제효소는 pH 5.0~9.0 사이에서 안정하였고, 5$0^{\circ}C$, 3시간 및 6$0^{\circ}C$, 30분까지는 비교적 안정하였다. 금속염 및 화학물질의 영향을 조사한 결과 Fe$^{2+}$, Ag$^{1+}$ 및 단백질변성제인 Hg$^{2+}$ 이온에 의해 효소활성이 크게 저해되었고, p-CMB, iodoacetic acid, urea, 2,4-DNP 및 EDTA에 의해 효소활성이 약간 저해되었다. 기질특이성을 조사한 결과 colloidal chitin 및 shrimp shell 유래의 chitin은 분해가능하였으나 crab shell 유래의 chitin, chitosan등은 분해하지 못하였다. Colloidal chitin에 대한 본 효소의 Km값은 0.11%였고, 분해율은 24시간 반응시 34%였다.

• Journal of Microbiology and Biotechnology
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• 제6권6호
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• pp.439-444
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• 1996

A bacterial strain, designated as WY22, producing extracellular chitinase was isolated from the soil around the Youngduck area, after enrichment culture in a medium containing $1{\%}$ (w/v) wet colloidal chitin as a sole carbon source. The isolate was identified as a strain of Bacillus sp. based on its morphological and physiological characteristics. It was observed that Bacillus sp. WY22 could inhibit the growth of Fusarium oxysporum with hyphal extention-inhibition assay on potato dextrose agar plate supplemented with $1{\%}$ collidal chitin. Optimum culture conditions of Bacillus sp. WY22 were examined for chitinase production in a chitin medium. High level production of chitinase was observed not only in the chitin medium but in a medium supplemented with $1{\%}$ N-glucosamine or lactose instead of chitin. The optimum concentrations of colloidal chitin and yeast extract were 3.0 and $0.5{\%}$, and the optimum culture conditions for initial pH of medium and temperature were 7.0 and $30^{\circ}C$, respectively, for the production of chitinase.

• Journal of Microbiology and Biotechnology
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• 제9권6호
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• pp.839-846
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• 1999

An extracellular chitinase-producing bacterial strain induced by colloidal chitin was isolated from sea sand and was identified to be a member of the genus Cytophaga. The chitinase was purified successively by 30-60% ammonium sulfate fractionation, and DEAE-Bio gel A column, Octyl-Sepharose CL-4B column, and DEAE-Bio gel A column chromatographies. The enzyme had a molecular mass of 59.75 kDa, and the amino terminal amino acid sequence was ATPNAPVISW MPTDXXLQNXS. The enzyme acted better on colloidal chitin as a substrate than on chitosan. For colloidal chitin and chitosan (Degree of Acetylation, 15-25%), $K_{cat}$ values were 0.60U/mg and 0.08U/mg, respectively. HPLC analysis of the enzymatic reaction products showed that the chitinase produced mostly N-acetyl-D-glucosarnine and di-N-acetylchitobiose. The optimum temperature and pH for the enzyme were $50^{\circ}C$ and 4.0, respectively. N-Bromosuccinimide and $Hg^{2+}$ inhibited the chitinase activity as much as 90%, and $Sb^{3+}$, diethylpyrocarbonate, and $Ag^{+}$ inhibited it by 50-70%.

• KSBB Journal
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• 제11권6호
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• pp.696-703
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• 1996

S. marcescens QM 81466 균주는 chitin 분해 효소(1mg/Lmedium)를 선택적으로 높게 생성시킬 수 있는 균주로서, chitin을 N-acetyl-$\beta$-D-glucosa­m mine(NAG)으로 효소적 가수분해를 할 때 chitinase와 chitobiase의 두 가지 가수분해 효소계를 구 성시킨다. 본 연구에서는 이 균주의 chitinase/chitobiase 생성을 위한 chitin 입자크기에 대한 최적화와, 회분 발효계에서 이 균주의 세포 밀도 배양에 따른 두 효소 생성의 변화를 조사하여 NAG 생산성의 증대를 시도하였다. 아울러. chitin과 CM­ chitin이 chitinase/chito biase 생성비 와 NAG 생성 에 미치는 영향을 검토하였는데, CM-chitin을 colloidal 및 결정성 chitin 대신에 사용했을 때, chitinase 활성을 약 7~10U/mL 증가시켰다. 이 경우에 있어서, chitinase/chitobiase의 비는 9:1로 서 NAG의 생성량이 3.0g/L로서 높게 나타났다.

• 한국미생물·생명공학회지
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• 제18권6호
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• pp.599-606
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• 1990

근해 연안 토양으로부터 Chitinase 활성이 우수한 균주를 분리하여 Aeromonas salmonicida로 동정하였으며, 분리균주의 효소생산 최적조건은 colloidal chitin 1.26, tryptone 2.95, $MgSO_4-7H_20$0.15, $K_2HP0_4$, 0.15, pH8.5, 27'C에서 48시간 진탕배양하였을 때였다. 효소의 정제는 배양액으로부터 ammonium sulfate 침전, affinity adsorption, hydroxylapatite chromatography, gel filtration을 통해 수율 29.7, 정제도 18.5배의 정제효소를 얻었다. 정제된 chitinase의 최적온도와 pH는 $50^{\circ}C$와 7.0이었고 pH 안정성은 pH5.0-9.0 사이였고 $50^{\circ}C$까지 안정하였으며 Km값은 1.276mg/ml, 분자량은 200,000 daltons으로 확인되었다.

• KSBB Journal
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• 제16권4호
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• pp.365-369
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• 2001

Serratia marcescens KY와 Serratia marcescens ATCC 27117 두 균주 모두 기본 배지에 $K_2$HPO$_4$ 농도를 0.2 g/L 농도 첨가할 경우 최대 chitinase 생성을 나타냈다. 그러나 세포 성장에 따른 균체량은 기본 배지에 $K_2$PHO$_4$ 농도 0.2 g/L 이상에서는 균체량은 약간 감소한다. 1.2% colloidal chitin과 0.2 g/L의 $K_2$PHO$_4$가 포함된 기본 배지에 MgSO$_4$를 0.2-0.25 g/L를 첨가하였을 때 최대의 chitinase 생성을 보이고, 두 균주 모두 K, P, Mg 및 기타 mineral을 영양 요구 인자로서 필요한다. Colloidal chitin을 1.2% 함유하고 있는 기본 배지에 각종 탄소원의 종류에 따른 세포 성장과 chitinase 생성은 colloidal chitin만을 첨가하였을 때가 상대적으로 가장 우수하고, 탄소원을 첨가할 경우 Serratia marcescens는 모든 탄소원에서 chitinase 생성이 억제되었다. 또한 질소원에 따른 세포 성장과 chitinase 생성은 Serratia marcescens KY와 Serratia marcescens ATCC 27117 모두 tryptone이 가장 우수하였고, 2.0 g/L의 질소원 농도까지는 질소원 농도가 증가함에 따라 chitinase 생성은 증가하다가 2.0 g/L 이상의 농도에서는 질소원 농도가 증가함에 따라 chitinase 생성은 감소하였다. 이들 질소원 중 chitinase 생성은 trypotone>yeast extract > beef extract > asparagine 순서로 chitinaserk 생성되므로, Serratia marcescens는 chitinase 생성에 있어 vitamin B군과 같은 질소원을 growth factor로 요구한다.

• 한국토양비료학회지
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• 제36권4호
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• pp.247-255
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• 2003

해안가 토양으로부터 강력한 chitinase 활성을 가진 Bacillus cereus KJA-118이 분리.동정되었다. B. cereus KJA-118은 1% colloidal chitin이 포함된 배지를 pH 6.0으로 조절한 후 $30^{\circ}C$ 에서 4일동안 호기적으로 배양했을 때 가장 높은 chitinase 효소활성을 보였다. 탄소원 (crab shell powder, chitin powder, colloidal chitin, and R. solani 균사)에 따른 chitinase 활성은 R. solani 균사를 사용했을 때 가장 높았다. Glycol chitin 0.01%가 포함된 gel에서 전기영동 후 활성 염색한 결과, B. cereus KJA-118에 의해 생산되는 chitinase는 분자량이 68, 47, 37 KDa인 3개의 isoform이 검출되었다. 액체 배지에서 미리 배양한 B. cereus KJA-118과 R. solani를 다시 혼합 배양했을 때, 곰팡이의 세포벽이 완전히 파괴되었다. R. solan가 감염된 토양에 B. cereus KJA-118의 배양액을 처리했을 때 28.1%의 오이 모잘록병 억제 효과를 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제5권2호
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• pp.80-86
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• 1995

The bacterial strain WC-17 able to produce chitinase was isolated from soil using an enrichment technique. The isolated strain was identified as Acinetobacter sp. judging by their morphological and physiological characterisitics. The optimal culture conditions for the production of chitinase of Acinetobacter sp. WC -17 are 1.5% colloidal chitin and 1 % tryptone at $30^{\circ}C$ with pH 6.5. Since the enzyme was rapidly produced in a culture supplied with chitin, glucose, or N-acetylglucosamine but not with other polymers and monosaccharide, the enzyme was considered to be an inducible enzyme. Notably N- acetylglucosamine and glucose were found to be effective inducers at low concentrations but repressors at excessive concentrations. The cultural supernatant of Acinetobacter sp. WC-17 inhibited the growth of phytopathogenic fungi such as P.oryzae, R.solani, and F.solani. Among the phytopathogenic fungi tested, P.oryzae was the most sensitive. The conventional agar plate (PDA containing 1 % colloidal chitin) method also produced the same result.