한밭 수목원의 과일과 꽃으로부터 효모의 분리

현세희,민진홍,김선아,이종수,김하근 한국균학회 2014 韓國菌學會誌 Vol.42 No.2

Yeasts are common inhabitants of the phyllosphere, but our knowledge of their diversity in various fruits andblossoms are limited. We collected different kinds of blossoms and unripened fruits from Hanbat arboretum, Daejeon, Korea atthe year of 2013. Yeasts were isolated by plating of suspensions prepared for collected samples onto YPD medium containingantibiotics. BLAST searches were subsequently performed for the comparison of the partially determined sequences of D1/D2domain of 26S rDNA. As a result, we isolated 57 yeast strains of 31 species from 29 different kinds of blossoms and 6 kinds offruits samples. We found huge differences in yeast flora depending on the sample collection season. 효모는 엽권의 중요한 서식자이지만, 효모의 분포와 역할을 정확하게 이해하고 있지 못한 형편이다. 2013년 봄과 가을에, 대전시 서구 만년동에 위치한 한밭 수목원에 식재되어 있는 각종 꽃과 과일로부터 효모를 분리하였다. 분리된효모 집락을 PCR 방법에 의해 26S rDNA의 D1/D2 지역을증폭하고 염기서열을 결정한 후, BLAST를 이용하여 데이터베이스와 비교하여 효모를 동정하였다. 그 결과 29종의꽃과 6종의 과일로부터, 14속 31종에 속하는 효모를 57균주를 분리하였다. 그 결과 봄과 가을, 계절에 따라서 같은수목원에서 분리되는 효모 종에 현저한 차이가 있는 것을발견하였다.

Characteristics of Two Unrecorded Yeasts from Wild Flowers in Ulleungdo, Korea

현세희,민진홍,이향범,김하근,이종수 한국균학회 2014 韓國菌學會誌 Vol.42 No.2

Two unrecorded yeasts, Meyerozyma caribbica UL5-1 and Pichia silvicola UL6-1 were screened from 58 yeasts whichwere isolated from wild flowers in Ulleungdo in Gyeongsangbuk-do, Korea. The morphological and cultural characteristics ofthese unrecorded yeasts were investigated. Both yeasts were oval in shape and formed pseudomycelia. P. silvicola UL6-1 formedascospore, but M. UL5-1 did not. P. silvicola UL6-1 and M. caribbica UL5-1 also grew in vitamin-free medium and 5% NaClcontainingyeast extract-peptone-dextrose medium. The two unrecorded yeasts assimilated glucose, galactose, xylose, cellobiose,trehalose, glycerol and sorbitol, and also fermented glucose, fructose and mannose. The supernatant of both M. caribbica UL5-1and P. silvicola UL6-1 showed high antihypertensive angiotensin I-converting enzyme inhibitory activity of 84.2% and 82.6%,respectively. Cell-free extract of P. silvicola UL6-1 also showed very high anti-diabetic α-glucosidase inhibitory activity (85.8%).

배 유과 가공제품의 품질 특성 및 저장 중 생균수 변화

민진홍, 현세희1, 나광출, 이종수, 김하근 배재대학교 자연과학연구소 2012 自然科學論文集 Vol.23 No.1

To develop the functional pear fruitlet product, we prepared dried pear fruitlet and fermented pear fruitlet, respectively. Then we investigated their general characteristics, and viable cell number changes during the storage at room temperature. We also determined preference for their tastes. Dried pear fruitlet contained more ash and crude proteins as well as minerals than fermented pear fruitlet. However fermented pear fruitlet contained higher amount of edible fiber than dried pear fruitlet. Fungi were not detected from dried pear fruitlet during the storage experiment at room temperature. Numbers of bacteria from dried pear fruitlet were increased from 1.1☓105/g at the beginning to2.6☓106/g at the last day of experiment. When we determined change of fungi number during the storage of fermented pear fruitlet, we cannot detect fungi at the beginning but we detected 4.4☓102/g of fungi at the last day of storage experiment. Bacteria from fermented pear fruitlet were detected at around 1.0☓109/g during the storage periods. Preference for fermented pear fruitlet was superior to dried one.

보문 : 전라북도 고군산열도의 선유도 일대 야생화들로부터 효모의 분리 및 생리기능성 현세희,한상민,이종수

현세희 ( Se Hee Hyun ),한상민 ( Sang Min Han ),이종수 ( Jong Soo Lee ) 한국균학회 2014 韓國菌學會誌 Vol.42 No.3

Sixty one yeast strains of 21 species were isolated from wild flowers in Gogunsanyeoldo, including Seonyudo of Jeollabuk-do, Korea, and identified by comparison of nucleotide sequences for PCR-amplified D1/D2 region of 26S rDNA using BLAST. Among them, Cryptococcus sp. including C. aureus SY1-4 were found to be dominant, and Metschnikowia sp. including M. reukaufii SY20-1 and Rhodotnula sp. such as R. ingeniosa SY1-1 were also abundantly isolated. Some physiological functionalities of the culture broth and cell-free extracts from 61 yeast strains were determined. Supernatant from Metschnilowia reukaufii SY44-6 showed anti-gout xanthine oxidase inhibitory activity of 49.6% and whitening tyrosinase inhibitory activity of 38.4%, respectively.

단보 : 국내 야생화들로부터 국내 미기록 효모, Rhodosporidium fluviale, Rhodosporidium paludigenum과 Candida sp. 80-J-3, Kluyveromyces thermotolerans의 특성

현세희 ( Se Hee Hyun ),이향범 ( Hyang Burm Lee ),이종수 ( Jong Soo Lee ) 한국균학회 2013 韓國菌學會誌 Vol.41 No.3

충남 금산군과 전남 완도군, 제주도 송악산 등지에서 서식하고 있는 야생화들로부터 Rhodosporidium fluviale 201-C-1과 Kluyveromyces thermotolerans 150-JE-2(1), Rodos-poridium paldigenum과 ㅊ무양ㅁ sp. 80-J-3등의 국내 미기록 효모들을 분리, 선발하여 이들의 미생물학적 특성들을 조사하였다. Rhodosporidium fluviale 201-C-1은 내염성의 무포자 효모로 의균사를 형성하였고 urease 활성을 갖고 있었다. Kluyveromyces thermotolerans 150-JE-2(1)는 내당성과 내염성을 가진 무포자효모로서 30℃, pH 4.0~6.0에서 잘 생육하였다. Candida sp. 80-J-3과 Rhodosporidium palu-digenum 86-J-1 균주들은 모두 무성생식을 ㅎ는 무포자 효모들이었고 YPD배지, Vitamin-free 배지와 5% NaCI을 함유한 YPD 배지 등에서 잘 생육하였다. 또한 Candida sp. 80-J-3 균주의 urease 활성은 음성이었으나 Rhodosporidium paludigenum 86-J-1 균주는 urease 활성이 양성이면서 pH 7.0~8.0에서 잘 생육하였다. Rhodosporidium fluviale 201-C-1 and Kluyveromyces thermotolerans 150-JE-2(1) were isolated from wild flowers in Geumsan-gun, Chungcheongnam-do and Wando-gun, Jeollanam-do, respectively. Rhodosporidium Paludigenum 86-J-1 and Candida sp. 80-J-3 were also isolated from flowers in Jeju island, Korea. Characteristics for these yeasts were not reported before. Therefore, morphological and physiological characteristics for these yeasts were investigated in this study. R. fluviale 201-C-1 was halophilic asporogenous yeast and formed Pseudomycelium. It also grew at 30°C and pH 4.0-6.0. K. thermotolerans 150-JE-2(1) was sugar-tolerant and halotolerant asporogenous yeast. It formed pseudomycelium and showed positive urease activity. Candida sp. 80-J-3 and R. Paludigenum 86-J-1 were asporogenous yeasts, and grew well in yeast extract-peptone dextrose(YPD) medium, potato-dextrose medium as well as vitamin-free medium and YPD medium containing 5% NaCI.

우리나라 해안 근접지역과 일부 내륙지역 야생화들로부터 분리한 국내 미기록 효모들의 특성

현세희 ( Se Hee Hyun ),이향범 ( Hyang Burm Lee ),김창무 ( Chang Mu Kim ),이종수 ( Jong Soo Lee ) 한국균학회 2013 韓國菌學會誌 Vol.41 No.2

Fourteen new records of yeasts were screened from one hundred thirty-three yeasts strains which were isolated from wild flowers in east, west and south coast near areas and inland areas, Korea. Candida silvae, Cryptococcus terrestris and Auriculibuller fuscus were newly isolated from wild flowers collected at Donghae and Andong, respectively. Candida quercitrusa, Hanseniaspora opuntiae, Sporobolomyces carnicolor, Candida saopaulonensis and Sporobolomyces phaffii were newly isolated from wild flowers in Geumsan-gun and Taean-gun in Chungcheongnam-do, respectively. Candida oleophila and Jaminaea angkoriensis were newly identified from flowers in Yesan-gun and Hannaella zeae was also newly identified from flowers of Daecheon. Hanseniaspora vineae and two other yeast strains including Pseudozyma hubeiensis were newly isolated and identified from flowers collected at Gwangju and Wando-gun, respectively. Morphological and physiological characteristics of their new recorded yeasts were investigated.

전라북도 선유도일대 야생화에서 분리한 국내 미기록 효모들의 특성 및 생리기능성

현세희 ( Se Hee Hyun ),한상민 ( Sang Min Han ),이종수 ( Jong Soo Lee ) 한국미생물생명공학회(구 한국산업미생물학회) 2014 한국미생물·생명공학회지 Vol.42 No.4

본 연구에서는 아직까지 우리나라 미생물관련 학술지에 보고되어 있지 않은 Rhodosporidium diobovatum SY4-2, Cryptococcus bestiolae SY7-1, Kazachstania unispora SY14-1, Kazachstania servazzii SY14-3, Pichia holstii SY20-2와 Cryptococcus tephrensis SY26-1 등 6종의 새로운 국내 미기록 효모들을 전라북도 고군산열도의 선유도 일대 야생화들에서 분리, 동정된 61균주에서 선별하였다. 이들 균주들 모두 구형과 난형으로 50% 포도당을 함유한 YPD 배지에서 잘 생육하였고, Pichia holstii SY20-2는 20% NaCl을 함유한 YPD 배지에서 생육하는 극도 호염성 효모이었다. Cryptococcus tephrensis SY26-1은 포도당을 자화시키지 못하였으나 Cryptococcus bestiolae SY7-1과 함께 유당을 자화시켰다. 6종의 미기록 효모 균주들의 생리기능성을 조사한 결과 이들 균주들의 무세포추출물들의 항당뇨성 α-glucosidase 저해활성이 86.8-98.6%로 매우 높았고, 특히 Kazachstania servazzii SY14-3의 무세포추출물이 98.6%로 가장 높았다. 또한, 미기록 효모 균주들의 배양상등액의 항고혈압성 안지오텐신 전환효소 저해활성도 58.6-80.4%로 비교적 높았다. Six kinds of newly recorded yeasts such as Rhodosporidium diobovatum SY4-2, Cryptococcus bestiolae SY7-1, Kazachstania unispora SY14-1, Kazachstania servazzii SY14-3, Pichia holstii SY20-2 and Cryptococcus tephrensis SY26-1 were screened from sixty one yeasts derived from wild flowers found in Seonyudo, Gogunsanyeoldo, Jeollabuk-do, Korea. All of them grew in 50% glucose-containing yeast extract-peptone-dextrose (YPD) broth and Pichia holstii SY20-2 was also halophile, growing in 20% NaCl-containing YPD broth. All of them, except Cryptococcus tephrensis SY26-1, were assimilated to glucose. Cell-free extract from Kazachstania servazzii SY14-3 showed the highest 98.6% of α-glucosidase inhibitory activity and maximal production of the α-glucosidase inhibitor was obtained with 24h incubation at 30oC. The antihypertensive angiotensin I-converting enzyme inhibitory activity of the unrecorded yeasts were showed 58.6-80.4% in their supernatants.

경기도와 제주도 야생화들로부터 효모의 분리, 동정 및 항통풍성 Xanthine oxidase 저해물질의 생산

현세희 ( Hyun Se Hee ),문혜연 ( Hye Yeon Mun ),이향범 ( Hyang Burm Lee ),김하근 ( Ha Kun Kim ),이종수 ( Jong Soo Lee ) 한국미생물생명공학회(구 한국산업미생물학회) 2013 한국미생물·생명공학회지 Vol.41 No.4

국내 야생화들로부터 효모를 분리하여 이들을 고부가가치의 의약이나 건강 식품산업에 응용하기 위한 연구의 일환으로 경기도와 제주도 일부 지역의 야생화들을 채집하여 효모들을 분리, 동정하고 이들 가운데 항통풍성 xanthine oxidase 저해물질 생산 균주를 선발한 후 생산 조건을 검토 하였다. 서울 구로동과 일부 경기도 지역 야생화 31점에서 26종, 54균주의 효모들을 분리, 동정하였고 이들 중 Pseudozyma sp.가 13종으로 가장 많이 분리되었다. 제주도 송악산과 성산 일출봉 지역의 야생화 21점에서 모두 14종, 32균주의 효모들을 분리, 동정하였고, 이들 균 중에서 Sporobolomyces ruberrimus가 6종으로 가장 많이 분리되었다. 86효모 균주들을 YEPD 배지에서 2일간 배양하여 각각의 배양 상등액과 무세포 추출물을 제조한 후 이들의 항통풍성 Xanthine oxidase 저해활성을 측정한 결과, Pseudozyma hubeiensis 228-S-1의 무세포 추출물이 19.5%의 가장 높은 Xanthine oxidase 저해활성을 보였다. 또한 Pseudozyma hubeiensis 228-S-1을 YEPD 배지에 접종하여 30oC에서 36시간 배양하였을 때, 세포내 항통풍성 xanthine oxidase 저해물질을 가장 많이 생산하였다. Several yeasts were isolated from flowers found in Gyonggi-do Province and Jeju island in Korea. They were then identified by a comparison of their PCR-amplified D1/D2 regions of 26S rDNA, internal transcribed spacer 1 and 2 inclusive of 5.8S rDNA, using the BLAST database. A total of fifty four yeast strains were isolated from wild flowers in Gyonggi-do and the genus Pseudozyma was noted as being dominant. A total of thirty two strains were isolated from Songaksan and Seongsan-ilchulbong in Jeju island and Sporobolomyces ruberrimus was seen to be dominant. The anti-gout xanthine oxidase inhibitory activities of the culture broths and cell-free extracts from eighty six yeast strains were then determined. The cell-free extracts of Pseudozyma hubeiensis 228-S-1 exhibited the highest xanthine oxidase inhibitory activity of 19.6%. The XOD inhibitor was also maximally produced when Pseudozyma hubeiensis 228-S-1 was cultured at 30oC for 36h in YEPD medium.

보문 : 경북 울릉도와 경남 욕지도 야생화들로부터 효모의 분리 및 종 분포 특성

현세희 ( Se Hee Hyun ),민진홍 ( Jin Hong Min ),이향범 ( Hyang Burm Lee ),김하근 ( Ha Kun Kim ),이종수 ( Jong Soo Lee ) 한국균학회 2014 韓國菌學會誌 Vol.42 No.1

Various yeasts from wild flowers of Ulleungdo in Gyeongsangbuk-do and Yokjido in Gyeongsangnam-do, Korea were isolated and identified by comparison of nucleotide sequences for PCR-amplified D1/D2 region of 26S rDNA using BLAST. Fortyeight yeast strains of twenty two species and sixty yeast strains of twenty five species were isolated from wild flowers of Ulleungdo and Yokjido, respectively. Only seven species were overlapped from the two different islands areas: Cryptococcus albidus, Cryptococcus laurentii, Metschnikowia reukafii, Pichia scolyti, Rhodotorula glutinis, Rhodotorula graminis and Rhodotorula mucilaginosa. Among forty species from two different islands, other thirty three species were restricted to specific collection site suggesting that each area has distinctive yeast flora.

총설 : 한국의 야생화에서 분리한 효모의 종 다양성과 생리기능성

현세희 ( Sehee Hyun ),한상민 ( Sangmin Han ),김하근 ( Hakun Kim ),이종수 ( Jongsoo Lee ) 한국균학회 2015 韓國菌學會誌 Vol.43 No.3

우리나라 야생화들의 효모 종 다양성을 확립하고 이들을 고부가가치의 건강산업에 응용하기 위한 자료를 얻고자 한국의 제주도, 울릉도, 욕지도와 선유도 등 4개의 섬지역 산들과 계족산, 오서산, 백암산과 덕유산 등 4개의 내륙지역산에서 개화한 야생화들을 2012년에서 2014년 사이에 수집하여 효모 134종 289균주를 분리 동정하였다. 분리한 289균주 중에서 Cryptococcus속 균주들이 가장 많이 분리되었고 Cryptococcus aureus를 포함하는 23균주들이 섬과 내륙 산지역에서 공통적으로 분리 동정되었다. 또한 단일 균주로는 Metschnikowia reukaufii가 전체 균주 중 10.3%인 30균주로 가장 많이 분리되었다. 분리한 균주들을 yeast peptone dextrose (YPD) 배지에서 24시간 배양하여 각각의 배양상등액과 무세포추출물을 제조한 후 이들의 주요 생리기능성을 측정한 결과, Candida sp. 78-J-2의 배양상등액이 22.5%의 항산화활성, Metschnikowia reukaufii SY44-6의 배양상등액이 49.6%의 항통풍성 xanthine oxidase 저해활성과 38.4%의 미백성 tyrosinase 저해활성을 보였다. This study focused on isolation of wild yeasts from natural flowers and elucidation of yeast diversity. Wild yeasts were isolated from various flowers collected from mountains on the islands including Jejudo, Ulleungdo, Yokjido, and Seonyudo as well as inlands including Gyejoksan, Oseosan, Beakamsan, and Deogyusan in Korea. Isolated yeasts were identified by comparison of nucleotide sequences for polymerrase chain reaction-amplified D1/D2 region of 26S rDNA or internal transcribed spacer 1 and 2 including 5.8S rDNA using BLAST. 289 strains belonging to 134 yeast species were isolated. Cryptococcus genus strains were the most frequently isolated species among the identified yeasts. Metschnikowia reukaufii was also frequently isolated. Twenty three species including Cryptococcus aureus were overlapped between those of mountains on islands and inland. Physiological functionalities such as antioxidant activity, xanthine oxidase inhibitory activity, and tyrosinase inhibitory activity for the 289 identified yeast strains were investigated using their supernatant and cell-free extracts. The supernatants of Candida sp. 78-J-2 and Metschnikowia reukaufii SY44-6 showed antioxidant activity of 22.5%, and anti-gout xanthine oxidase inhibitory activity of 49.6%, respectively.