Metagenomic Lactic Acid Bacterial Diversity during Mulkimchi Fermentation Based on 16S rRNA Sequence

김바오로,서원택,Min Geun Kim,윤한대,Kye Man Cho 한국응용생명화학회 2012 Applied Biological Chemistry (Appl Biol Chem) Vol.55 No.6

Lactic acid bacterial diversity and the composition of individual bacterial communities during the fermentation of mulkimchi were examined using a polymerase chain recation (PCR)-based approach. Based on 16S rRNA sequence similarity values, a total of fifteen different lactic acid bacterial species were found in eight sampling sites, including Lactobacillus alimentarius,Lactobacillus brevis, Lactobacillus farciminis, Lactobacillus fabifermentans, Lactobacillus nantensis, Lactobacillus parabrevis,Lactobacillus plantarum, Lactobacillus versnoldensis, Lactobacillus zymae, and Lactobacillus sp., Leuconostoc pseudomesenteroides,Weissella cibaria, Weissella confusa, and Weissella sp. The prevalence of We. cibaria, belonging to the Weissella genus, was the highest (86.7%) at 0 h (initial stages) and gradually decreased at 72 h (rancid stage). In contrast, La. plantarum was observed at 36 h (16.7%, over-ripening stage) and gradually increased up to 84 h (70.0%, rancid stage) during mulkimchi fermentation. We. cibaria was found to be associated with the microorganisms that were present during the initial stage of fermentation, whereas La. plantarum was associated with the production of lactic acid in the over-ripening and rancid stages during fermentation at 30oC±2.

공액리놀레산 생성 Lactobacillus plantarum 선발 및 이를 이용한 콩-분말 두유에서 공액리놀레산 생산

김바오로,이병원,황정은,이유영,이춘우,김병주,박지영,심은영,모하메드 아지줄 하크만,이동훈,이진환,안민주,이희율,고종민,김현태,조계만,Kim, Baolo,Lee, Byong Won,Hwang, Chung Eun,Lee, Yu-Young,Lee, Choonwo,Kim, Byung Joo,Park, Ji-Yong,Sim, Eun-Yeong,Haque, Md. A 한국미생물학회 2015 미생물학회지 Vol.51 No.3

본 연구에서는 발효식품으로부터 16종의 공액리놀레산(CLA) 생성 젖산균을 분리하였다. 이들 균주 중, S48 및 P1201 균주는 다른 젖산균들보다 더 높은 CLA를 생성하였다. 두 균주는 형태학적, 생리학적, 화학적 및 분자유전학적 특징에 따라 Lactobacillus plantarum로 동정하였다. pH 2.0 인공위액산에서 4시간 배양 후 이들 균주의 생존율은 각각 59.57% 및 6.22%을 나타내었다. 이들 균주는 $37^{\circ}C$에서 48시간 동안 상이한 자유 리놀레산 함량이 함유되어 있는 8% 탈지분유 배지에서 cis-9, trans-11 및 trans-10, cis-12 CLA 이성체가 생성되었고, 두 CLA 이성체는 48시간 배양 동안 지속적으로 증가하였다. 발효 48시간 후, 증자콩-분말 두유의 CLA 함량은 생콩 및 볶음콩-분말 두유에서 보다 높은 생성능을 보였다. 특히, 증자콩-분말 두유의 CLA 함량은 각각 S48 및 P1201 균주에서 $183.57{\mu}g/ml$ 및 $198.72{\mu}g/ml$ 생성되었다. In this study, a total of 16 conjugated linoleic acid (CLA) producing lactic acid bacteria (LAB) were isolated from fermented foods. Among those strains, the S48 and P1201 strains were capable of producing higher CLA contents than other LABs. The two strains were classified as Lactobacillus plantarum based on morphological, physiological, chemotaxonomic, and molecular-genetic properties. The survival rates of these strain appeared to be 59.57% and 62.22% under artificial gastric conditions after 4 h at pH 2.5, respectively. These strains produced the cis-9, trans-11, and trans-10, cis-12 CLA isomers from 8% skim milk medium supplemented with the different free LA concentration at $37^{\circ}C$ for 48 h and the production of two CLA isomers constantly increased in the growth until 48 h of incubation. After 48 h of fermentation, the levels of CLA appeared highest in steamed soy-powder milk than fresh and roasted soy-powder milks. In particular, the CLA contents were produced $183.57{\mu}g/ml$ and $198.72{\mu}g/ml$ from steamed soy-powder milk after fermentation (48 h) with S48 and P1201 strains, respectively.

하느님께 드리는 편지 1

김 바오로 우리신학연구소 2009 갈라진 시대의 기쁜소식 Vol.- No.876

Microbial Diversity in the Enrichment Cultures from the Fermented Beverage of Plant Extract Using Ribosomal RNA Sequence Analysis

이총규,김바오로,강민영,이희율,황정은,안민주,서원택,조계만,Lee, Choung Kyu,Kim, Baolo,Kang, Young Min,Lee, Hee Yul,Hwang, Chung Eun,Ahn, Min Ju,Seo, Weon Taek,Cho, Kye Man The Microbiological Society of Korea 2014 미생물학회지 Vol.50 No.4

식물추출발효음료(fermented beverage of plant extract, FBPE)는 과일류, 야채류, 약초류, 및 해조류를 설탕에 의한 당장법으로 발효시킨 액상발효음료이다. 본 연구에서는 FBPE의 이화학적 특성과 16S 및 26S rRNA 염기서열을 이용하여 집식배양된 FBPE의 미생물 다양성을 조사하였다. FBPE의 pH는 3.48, 산도는 1.68%, 당도는 $70^{\circ}$, 환원당은 1,026 g/L 및 알코올은 3.5%이었다. 유리당과 유기산은 glucose (567.83 g/L)와 tartaric acid (936.8 mg/L)로 나타났다. Lactobacillus homohiochii는 모든 집식배양 시료에서 우점종이었고 L. fructivorans는 20B 라이브러리에서만 나타났다. 세 가지의 다른 당 농도에서 집식배양된 FPEB의 우점종은 0Y 라이브러리(설탕농도 0%)에서는 Candida zeylanides (45.2%), 20Y 라이브러리(설탕농도 20%)에서는 C. lactis-condensi (35.7%)와 C. zeylanoides (35.7%) 및 40Y 라이브러리(설탕농도 40%)에서는 C. lactis-condensi (38.1%)이었다. 이외에 0Y 라이브러리에서는 C. lactis-condensi (40.5%), Pichia farinosa (7.1%), C. parapsilosis(4.8%) 및 Zygosaccharomyces bailii (2.4%)가 나타났으며, 20Y 라이브러리에서는 P. guilliermondii (14.3%), Pichia farinosa (7.1%), C. parapsilosis (4.8%) 및 Kazachstania exigua (2.4%)로 나타났고 40Y 라이브러리에서는 C. zeylanoides (30.9%), C. parapsilosis (11.9%), Z. bailii (11.9%), Pichia farinosa (4.8%), P. guilliermondii (2.4%)이 확인되었다. 이 결과는 앞으로 FBPE의 미생물 변화 연구에 아주 유용한 자료로 제공할 수 있다. A beverage was produced by the fermentation of mixed extracts from the various fruits, vegetables, algae, and medical herbs. The physicochemical properties of the fermented beverage of plant extracts (FBPE) and microbial diversity were analyzed in cultures enriched from FBPE using 16S and 26S rRNA gene sequence analyses. The pH, acidity, $^{\circ}brix$, reducing sugar, and alcohol contents of the FBPE were determined to be the 3.48, 1.68%, 70.0, 1,026 g/L, and 3.5%, respectively. The most abundant free sugar and organic acid in the FBPE were glucose (567.83 g/L) and tartaric acid (93.68 mg/L), respectively. Lactobacillus homohiochii was the predominant species in all enriched culture samples: 100% of the species in 0B (0% sugar) and 40B (40% sugar) libraries and 95.6% of 20B library (20% sugar). Lactobacillus fructivorans was detected in the 20B library. The predominant species in the samples of enrichment cultures collected from FBPE with three different sugar concentrations were: Candida zeylanoides (45.2%) in the 0Y library (0% sugar), Candida lactis-condensi (35.7%) and C. zeylanoides (35.7%) in the 20Y library (20% sugar), and C. lactis-condensi (38.1%) in the 40Y library (40% sugar). This result may provide a useful frame of reference for further analyses of microbial population dynamics in FBPE.

Rhizopus oryzae CCS01로 제조된 쌀누룩을 이용한 쌀-밀 막걸리의 품질 특성

서원택,조현국,이주영,김바오로,조계만,Seo, Weon-Taek,Cho, Hyeon-Kook,Lee, Ju-Young,Kim, Baolo,Cho, Kye-Man 한국미생물학회 2012 미생물학회지 Vol.48 No.2

본 연구에서는 시판누룩에서 분리한 Rhizopus oryzae CCS01로 쌀누룩을 제조하고 이를 이용하여 쌀-밀 막걸리 제조하였다. 쌀누룩은 시판누룩보다 ${\alpha}$-amylase 활성은 약 1.8-2.4배 정도 높게 나타났다. 쌀과 밀의 비율이 4:6일 때 알코올 생성량은 12.4% 있었으며, 전체적인 기호도면에서 가장 우수하였다. 시판누룩(산성, 진주 및 송학)으로 제조한 쌀-밀 막걸리의 pH는 발효가 진행됨에 따라 감소하였으나, 쌀누룩으로 제조한 쌀-밀 막걸리의 pH는 증가하였다. 한편 산도 및 당도, 알코올 함량은 발효가 진행됨에 따라 증가하였다. 특히 발효 종기 알코올 함량은 쌀누룩으로 제조한 쌀-밀 막걸리가 12.0%로 시판 누룩으로 제조한 막걸리보다 높게 나타났다. To improve of the quality of Korea traditional wheat-rice wine (makgeolli) production, we used a rice fermentation starter (rice nuruk) made by inoculation of Rhizopus oryzae CCS01 which was isolated and selected from commercial nuruk. Amylase activity of a rice nuruk was 1.8-2.4 times higher than those of commercial nuruks. The best acceptability of wheat-rice wine in a sensory test was observed at 4 : 6 ratio of wheat-rice mash at experimental condition. During the fermentation period, pH of wheat-rice makgeolli made with a rice nuruk was higher compared to those made with commercial nuruks such as Sanseong, Jinju, and Songhak. Acidity of makgeolli mash was lower in case of using a rice nuruk and birx and alcohol production were higher compared to those of makgeolli mash using commercial nuruks. Highest alcohol production was observed at makgeolli mash using a rice nuruk and 12% of alcohol was produced at fermentation end. These results suggest that production of a new type of wheat-rice makgeolli using a rice nuruk was possible.

추출용매에 따른 산양삼의 진세노사이드(Rg1, Rb1) 함량과 radical-scavenging 활성 비교

김수철(Su Cheol Kim),황정은(Chung Eun Hwang),김바오로(Ba O Lo Kim),이기현(Ki Hyun Lee),이진환(Jin Hwan Lee),조계만(Kye Man Cho),주옥수(Ok Soo Joo) 한국식품저장유통학회 2021 한국식품저장유통학회지 Vol.28 No.2

ITS 염기서열 분석 및 CAPS를 이용한 조이시아 속(Zoysia) 들잔디와 갯잔디의 구별

홍민지(Min-Ji Hong),양대화(Dae-Hwa Yang),정옥철(Ok-Cheol Jeong),김양지(Yang-Ji Kim),박미영(Mi-Young Park),강홍규(Hong-Gyu Kang),선현진(Hyeon-Jin Sun),권용익(Yong-Ik Kwon),박신영(Shin-Young Park),양바오로(Paul Yang),송필순(Pill-Soon So 한국원예학회 2017 원예과학기술지 Vol.35 No.3

Zoysia 속 잔디는 학교운동장 및 공원, 골프장, 스포츠경기장과 같이 다양한 장소에 식재되고 있는 중요한 잔디이다. 해안가에서 자생하는 Zoysia 속 들잔디와 갯잔디는 외부 형태적 특성이 유사하여 외부 형태적 분류 뿐 만 아니라 분자생물학적 분류도 필요하다. 본 연구에서는 nrDNA ̵ ITS(Internal Transcribed Spacer)의 DNA 바코드 분석을 통해서 자생하는 들잔디와 갯잔디의 분자생물학적 신속한 분류체계를 확립하고자 하였다. 이를 위해 난지형 잔디인 Zoysia 속 들잔디(Z. japonica) 및 갯잔디(Z. sinica)와 한지형 대표 잔디인 크리핑 벤트그라스(A. stolonifera) 및 켄터키 블루그라스(P. pratensis)의 nrDNA - ITS 염기서열을 확보하였다. 확보된 들잔디및 갯잔디, 크리핑 벤트그라스, 켄터키 블루그라스의 ITS 염기서열 전체 구간은 각 686bp와 687bp, 683bp, 681bp으로 확인되었으며, nrDNA - ITS 내부 염기서열구간 분석 결과, ITS1의 크기는 248 ̵ 249bp, ITS2는 270 - 274bp, 5.8S rDNA는 163 - 164bp의 차이로, 각 4종의 잔디가 ITS 염기서열을 이용하여 식별되었다. 특히, 들잔디와 갯잔디 nrDNA-ITS 염기서열은 19 염기(2.8%) 차이를 나타냈으며, ITS1과 ITS2의 G + C 함량은 55.4 ̵ 63.3% 임을 확인하였다. 이러한 들잔디와 갯잔디의 ITS 염기서열 차이를 바탕으로 CAPS 마커로 전환하여 대조구 및 수집된 자생 Zoysia 속 잔디 영양체 62개체를 분석한 결과, 외부형태학적 분류법으로 들잔디 개체, 갯잔디 개체로 동정되었지만, ITS CAPS 마커를 이용한 분자생물학적 분류법으로 들잔디 36개체와 갯잔디 22개체 뿐만 아니라 들잔디와 갯잔디간의 자연교배종 4개체도 식별하였다. 이상의 결과에서 들잔디와 갯잔디는 ITS 염기서열 및 ITS 기반 CAPS를 통하여 식별할 수 있을 것으로 판단된다. Zoysiagrasses are important turf plants used for school playgrounds, parks, golf courses, and sports fields. The two most popular zoysiagrass species are Zoysia japonica and Zoysia sinica. These are widely distributed across different growing zones and are morphologically distinguishable from each other; however, it is phenotypically difficult to differentiate those that grow along the coastal line from those in beach area habitats. A combination of morphological and molecular approaches is desirable to efficiently identify these two plant cultivars. In this study, we used a rapid identification system based on DNA barcoding of the nrDNA-internal transcribed spacer (ITS) regions. The nrDNA-ITS regions of ITS1, 5.8S nrDNA, and ITS2 from Z. japonica , Z. sinica , Agrostis stolonifera , and Poa pratensis were DNA barcoded to classify these grasses according to their molecular identities. The nrDNA-ITS sequences of these species were found at 686 bp, 687 bp, 683 bp, and 681 bp, respectively. The size of ITS1 ranged from 248 to 249 bp, while ITS2 ranged from 270 to 274 bp. The 5.8S coding region ranged from 163 - 164bp. Between Z. japonica and Z. sinica , nineteen (2.8%) nucleotide sites were variable, and the G+C content of the ITS region ranged from 55.4 to 63.3%. Substitutions and insert/deletion (indel) sites in the nrDNA-ITS sequence of Z. japonica and Z. sinica were converted to cleaved amplified polymorphic sequence (CAPS) markers, and applied to the Zoysia grasses sampled to verify the presence of these markers. Among the 62 control and collected grass samples, we classified three groups: 36 Z. japonica , 22 Z. sinica , and 4 Z. japonica /Z. sinica hybrids. Morphological classification revealed only two groups; Z. japonica and Z. sinica . Our results suggest that used of the nrDNA-ITS barcode region and CAPS markers can be used to distinguish between Z. japonica and Z. sinica at the species level.