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산화적 스트레스에 대한 Bacillus subtilis의 thiol peroxidase 유전자의 생리적인 기능
김하근,김성진,Kim, Ha-Kun,Kim, Sung-Jin 배재대학교 자연과학연구소 2005 自然科學論文集 Vol.15 No.1
산화적 스트레스에 대한 Bacillus subtilis의 thiol peroxidase 유전자의 생리적인 기능을 연구하기 위해 thiol peroxidase 유전자의 기능이 손상된 녹아웃 돌연변이주를 상동성 재조합에 의해 제조하였다. 호기적 조건에서 배양할 때 야생형과 녹아웃 돌연변이주 사이에는 성장속도에서 차이를 관찰할 수 없었다. 그러나 paraquat 처리할 때와는 달리 $H_2O_2$와 cumene hydroperoxide (CHP)에 의한 산화적 스트레스에 대해 역할을 하고 있음을 시사하는 결과이다. In Order to investigate the physiological role of thiol peroxidase in Bacillus subtilis, a thiol peroxidase (btpx) knock-out mutant was generated by homologous recombination. The growth of btpx knock-out mutant in aerobic condition showed a similar pattern with that of wild type of Bacillus subtilis 168/ But btpx knock-out mutant showed a retarded growht in response to oxidative stress such as $H_2O_2$, cumene hydroperoxide (CHP) treatments.
대장균에서 발현되는 Cellulomonas fimi $\beta$-glucosidase의 효소학적 특징
김하근,Kim, Ha-Kun 배재대학교 자연과학연구소 1996 自然科學論文集 Vol.8 No.2
Cellulomonas fimi에서 유래한 $\beta$-glucosidase 유전자를 갖고 있는 대장균으로부터 $\beta$-glucosidase 효소를 정제하였다. 전기 영동과 크로마토그라피 실험을 수행함으로써 정제된 효소의 분자량은 56,000 달톤이며 단일 폴리펩티드로 구성되어 있음을 알 수 있었다. 정제된 $\beta$-glucosidase 효소는 당이 $\beta$-결합을 하고 있는 cellobiose, PNPG, PNPC 등의 기질에 대하여 작용하여 분해시킬 수 있었으나, $\alpha$-결합을 갖고있는 maltose는 분해할 수 없었으므로, $\beta$-결합에 대한 기질 특이성을 갖고 있음을 알았다. 철, 수은, 구리 등의 중금속 이온들에 의해 효소 활성이 저해되었고 DTT에 의해 효소의 활성이 활성화됨을 보임으로써 $\beta$-glucosidase 효소의 활성화 부위는 -SH 기가 중요하게 작용하고 있음을 시사하였다. The $\beta$-glucosidase enzyme was purified from E. coli carrying Cellulomonas fimi $\beta$-glucosidase gene. SDS-PAGE and analytical gel filtration revealed that molecular weight of this enzyme was 56,000 dalton and consisted of a single polypeptide.Inhibition caused by heavy metals and activation by dithiothreitol suggest the existence of essential thiol group in the enzyme. The enzyme was not active on maltose (glucose $\alpha$-1,4-glucose) which has a $\alpha$-linkage, whereas it was active on lactose (glucose $\beta$-1,4-glucose), PNPG (p-nitrophenyl $\beta$-D-glucopyranoside) and PNPC (p-nitrophenyl $\beta$-D-cellobioside), although its reaction rates were different.
동해안 하조대와 경포대 해수욕장 주변 토양으로부터 분리한 국내 미기록 야생효모들의 균학적 특성과 생리활성
김하근 ( Ha-kun Kim ),김지윤 ( Ji-yoon Kim ),한상민 ( Sang-min Han ),김창무 ( Changmu Kim ),이종수 ( Jong-soo Lee ) 한국균학회 2019 韓國菌學會誌 Vol.47 No.3
The goal of this study was to isolate and characterize the wild yeast strains in the soils of Hajodae and Gyungpodae Beaches, Gyungpo Lake in Kangwon-do, Korea and evaluate their anti-demential and tyrosinase inhibitory activities. Among the 27 yeast strains isolated from 30 soil samples from Hajodae Beach, Cyberlindnera culbertsonii HJ31-1, Dothichiza pithyophila HJ22-4, Metschnikowia bicuspidata HJ16-1, and Sakaguchia cladiensis HJ14-1 were recorded for the first time in Korea. Additionally, six of the 55 wild yeast strains isolated from 45 soil samples from Gyungpodae Beach and Gyungpo Lake had not previously been detected in Korea, such as Aureobasidium melanogenum JR3-1. The microbiological characteristics and anti-demential and tyrosinase inhibitory activities of these previously unrecorded yeasts were investigated. Almost all of them were oval-shaped and had ascospores except for Dothichiza pithyophila HJ22-4, Diutina siamensis JR37-5, and Canadida gelsemii DC 35-1. Furthermore, they were all sugar-tolerant and able to grow in 20% glucose-containing YPD broth, except for Cyberlindnera culbertsonii HJ31-1. Diutina siamensis JR37-5 had the highest tyrosinase inhibitory activity (30.9%).
한상민,이상엽,김하근,이종수,Han, Sang-Min,Lee, Sang-Yeop,Kim, Ha-Kun,Lee, Jong-Soo 한국균학회 2017 韓國菌學會誌 Vol.45 No.1
충청남도와 전라북도에 걸쳐있는 대둔산과 충청남도 청양군 칠갑산의 각종 나뭇잎 87점을 수집하여 야생효모들을 분리, 동정하였다. 대둔산 나뭇잎들에서는 Cryptococcus tephrensis JSL0131를 포함하는 Cryptococcus속 균이 18균주로 가장 많이 분리되었고 Aureobasidium pullulans도 16균주가 분리되었다. 이들 중에서 Asterotremella humicola JSL502, Barnettozyma californica JSL503, Bullera unica JSL501, Bulleromyces albus JSL505와 Candida vaccinia JSL507이 국내 희귀 효모들이었고 Bulleromyces albus JSL505와 Bullera unica JSL501은 10%의 NaCl을 함유한 배지에서도 생육하는 호염성 효모였다. 칠갑산 나뭇잎에서는 Pseudozyma aphidis JSL614를 포함하는 Pseudozyma속 균이 가장 많이 분리 되었다. Rhodotorula phylloplana JSL515, Rhodotorula pinicola JSL516과 Schwanniomyces vanrijiae JSL517과 Sporisorium loudetiae JSL518 등 4균주들이 국내 희귀 종이었고 Sporisorium loudetiae JSL518은 포자와 의균사를 형성하였으며 Rhodotorula pinicola JSL516과 Schwanniomyces vanrijiae JSL517은 5% NaCl을 함유한 YPD 배지에서 생육하는 호염성 효모이었다. Several yeast strains were isolated from leaves collected from Mt. Daedun and Mt. Chilgap in Korea. A total of 79 strains of 32 species were isolated from 44 leaves obtained from Mt. Daedun. Cryptococcus spp., including Cryptococcus magnus, were found to be dominant, while 16 strains of Aureobasidium pullulans were isolated. Among the strains isolated from Mt. Daedun, Asterotremella humicola JSL502, Barnettozyma californica JSL503, Bullera unica JSL501, Bulleromyces albus JSL505, and Candida vaccinii JSL507 are considered rare yeast species in Korea. All of these rare strains, except for Bulleromyces albus JSL505, formed ascospores. Bullera unica JSL501 was able to grow in yeast extract-peptone-dextrose (YPD) medium containing 10% NaCl. Twenty-five yeast strains of 18 species were isolated from 43 leaves obtained from Mt. Chilgap. Six strains of Pseudozyma spp., including three strains of Pseudozyma aphidis, were isolated in the dormant state. Of these strains, the occurrence of Rhodotorula phylloplana JSL515, Rhodotorula pinicola JSL516, Schwanniomyces vanrijiae JSL517, and Sporisorium loudetiae JSL518 is rare in Korea. Only Sporisorium loudetiae JSL518 formed ascospores, while Rhodotorula pinicola JSL516 and Sporisorium loudetiae JSL518 formed pseudomycelia. The halophilic yeast Schwanniomyces vanrijiae JSL517 was isolated and observed to grow in YPD medium containing 5% NaCl.
낙동강과 영산강 담수와 주변 토양으로부터 야생효모의 분리 및 동정
한상민 ( Sang-min Han ),김하근 ( Ha-kun Kim ),이향범 ( Hyang-burm Lee ),이종수 ( Jong-soo Lee ) 한국균학회 2016 韓國菌學會誌 Vol.44 No.4
Diverse wild yeast were isolated from freshwaters and soils of Nakdong and Yeongsan rivers in Korea and identified by the comparison of polymerase chain reaction-amplified nucleotide sequences of the internal transcribed spacer region (including the 5.8S rRNA) and D1/D2 regions of 26S rDNA, using BLAST. In total, 15 strains belonging to 9 species were isolated from 25 samples, out of which Aureobasidium pullulans and Cryptococcus bestiolae were dominant. Candida ghanaensis JSF0127 and Meira geulakonigii JSF0130 were identified as unrecorded yeasts, for which their mycological characteristics were investigated. These unrecorded yeasts formed ascospores and grew in yeast extract peptone dextrose medium containing 5% NaCl.
시판 막걸리의 이화학적 특성과 젖산균 함량 및 생리기능성
김영헌 ( Young Hun Kim ),민진홍 ( Jin Hong Min ),강민구 ( Min Gu Kang ),김재호 ( Jae Ho Kim ),안병학 ( Byung Hak Ahn ),김하근 ( Ha Kun Kim ),이종수 ( Jong Soo Lee ) 한국미생물생명공학회(구 한국산업미생물학회) 2012 한국미생물·생명공학회지 Vol.40 No.4
본 연구는 우리나라 막걸리의 우수성을 발굴하고자 60종의 시판 생 막걸리와 저온살균 막걸리를 수집하여 이들의 주요 이화학적 성질과 젖산균 및 항고혈압활성 등의 생리기능성을 조사하였다. 생 막걸리의 에탄올 함량은 3.6%~9.6% 이 었고 이들의 잔당 함량은 0.20%에서 4.52%, 아미노태질소 함량은 3.50~101.9 mg%로 다양하였다. 저온살균 막걸리의 경우 에탄올 함량은 4.0%~7.0%로 생 막걸리와 비슷하였으나 잔당함량은 0.98%~7.57%로 생 막걸리보다 높았다. 또한 시판 생 막걸리의 젖산균 생균수는 평균 2.0×108 CFU/mL이었다. 여러가지 생리기능성 중 시판 EDS-14 생 막걸리와 PWR-12 저온살균 막걸리의 항고혈압성 안지오텐신 전환효소 저해활성이 각각 89.0%와 87.0%로 매우 높았으나 여타의 생리기능성은 30% 미만으로 낮았다. 또한 β-Glucan 함량은 생 막걸리와 저온살균 막걸리가 각각 건조고형물 g당평균 14.1%와 14.6%을 보였다. The goal of this study was to characterize the physicochemical properties, lactic acid bacteria and physiological functionality of Korean commercial Makgeolli. We collected 60 kinds of raw and pasteurized Makgeolli commercially available in Korea. Their physicochemical properties, lactic acid bacteria content and physiological functionalities were determined. In the commercial raw Makgeolli, ethanol and residual sugar content ranged from 3.6% to 9.6% and 0.20% to 4.52%, respectively. Furthermore, the raw Makgeolli contained 3.50 to 101.9 mg% of amino nitrogen. In the pasteurized Makgeolli, ethanol content ranged from 4.0% to 7.0% and their residual sugar content were 0.98% to 7.57%. The raw Makgeolli contained 2.0 ent3~1.2 ine8 CFU/mL of lactic acid bacteria. Among several functionalities, the antihypertensive angiotensin I-converting enzyme inhibitory activities of EDS-14 raw Makgeolli and PWR-12 pasteurized Makgeolli were very high, at 89.9% and 87.0% respectively. The other functionalities were seen to be below 30% or not detectable. The β-Glucan contents of the raw and pasteurized Makgeolli were noted as average, at 14.1% and 14.6%, respectively.
항고혈압성 안지오텐신 전환효소 저해제를 생산하는 새로운 효모의 선별 및 저해물질 최적 생산조건
강민구 ( Min Gu Kang ),김하근 ( Ha Kun Kim ),이성훈 ( Sung Hun Yi ),임성일 ( Sung Il Lim ),이종수 ( Jong Soo Lee ) 한국균학회 2011 韓國菌學會誌 Vol.39 No.3
Forty eight strains of yeast were cultured in potato dextorse(PD) broth at 30˚C for 24 hr and centrifuged with 12,000 rpm for 20 min. After concentrated the cultures, antihypertensive angiotensin I-converting enzyme(ACE) inhibitory activities of its concentrates were investigated. Among them, the concentrates from Saccharomyces cerevisiae Y183-3 showed the highest ACE inhibitory activity of 71.8%. The ACE inhibitor from Saccharomyces cerevisiae Y183-3 was maximally produced when Saccharomyces cerevisiae Y183-3 cultured in PD broth at 30˚C for 36 hr.
금강 중류 공주시 주변 물과 토양으로부터 야생효모의 분리 및 국내 미기록 효모 특성
한상민 ( Sang-min Han ),김지윤 ( Ji-yoon Kim ),이향범 ( Hyang Burm Lee ),김하근 ( Ha-kun Kim ),이종수 ( Jong-soo Lee ) 한국균학회 2018 韓國菌學會誌 Vol.46 No.2
The goal of this study was to elucidate wild yeast diversity of the midstream region of the Geumgang river, Gongju, Chungnam province, Korea. Fifty strains of 24 species of wild yeasts were isolated from 45 water and soil samples among which Cryptococcus spp. (11 strains), Cryptococcus magnus (7 strains), Rhodotorula spp. (9 strains), and Hanseniaspora spp. (6 strains) were dominant. Four species, Candida chauliodes WJSF 0201, Candida oleophila WJSF 0202, Candida catenulata WJSF 0203, and Candida jaroonii WJSF 0204, represented newly recorded yeasts in Korea. All of these unrecorded yeasts were oval in shape, formed ascospores and pseudomycelium, and grew in vitamin-free medium. Candida oleophila WJSF 0202 was thermophilic which can grow at 37°C.