Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Optimization of cultivation conditions for pullulan production from Aureobasidium pullulans MR by response surface methodology

반응표면분석법을 이용한 Aureobasidium pullulans MR의 풀루란 생산을 위한 배양 조건 최적화

  • Jo, Hye-Mi (Department of Food Science and Biotechnology, Kyung Hee University) ;
  • Kim, Ye-Jin (Department of Food Science and Biotechnology, Kyung Hee University) ;
  • Yoo, Sang-Ho (Department of Food Science & Biotechnology, Carbohydrate Bioproduct Research Center, Sejong University) ;
  • Kim, Chang-Mu (National Institute of Biological Resources) ;
  • Kim, KyeWon (Academic Industry Cooperation, Hankyong National University) ;
  • Park, Cheon-Seok (Department of Food Science and Biotechnology, Kyung Hee University)
  • 조혜미 (경희대학교 식품생명공학과) ;
  • 김예진 (경희대학교 식품생명공학과) ;
  • 유상호 (세종대학교 식품공학과) ;
  • 김창무 (국립생물자원관 미생물자원과) ;
  • 김계원 (국립한경대학교 산학협력단) ;
  • 박천석 (경희대학교 식품생명공학과)
  • Received : 2020.12.21
  • Accepted : 2021.02.08
  • Published : 2021.04.30
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Abstract

Aureobasidium pullulans, a black yeast, produces pullulan, a linear α-glucan composed of maltotriose repeating units linked by α(1→6)-glycosidic linkages. Pullulan can be widely used in food, cosmetic, and biotechnology industries. In this study, we isolated eight strains of A. pullulans from Forsythia koreana, Magnolia kobus DC., Spiraea prunifolia var. simpliciflora, Cornus officinalis, Cerasus, and Hippophae rhamnoides. Among them, A. pullulans MR was selected as the best pullulan producer. The effects of a carbon source, a nitrogen source, and pH on pullulan production were examined. The optimal cultivation conditions for pullulan production by A. pullulans MR were determined by response surface methodology as 15% sucrose, 0.4% soy peptone, and an initial pH of 7 at 26℃. Under these conditions, the predicted pullulan production was 47.6 g/L, which was very close to the experimental data (48.9 g/L).

본 연구에서는 A. pullulans MR의 풀루란 생산에 있어 배양 조건에 따른 변화를 살펴보았다. 먼저 시간과 온도에 따른 건조된 균체량과 풀루란 생산량의 변화를 비교하였으며, 기존의 AYS배 지내 탄소원과 혼합질소원 변화에 따른 풀루란 생산량을 비교하였다. 그 결과 sucrose와 ammonium sulfate, soy peptone을 본 실험의 최적 탄소원과 혼합질소원으로 결정하였으며, 그 후 반응표면분석법을 통해 탄소원인 sucrose와 질소원인 soy pepetone의 최적 농도와 최적 초기 pH를 알 수 있었다. 분산분석 결과를 토대로 비교하였을 때 A. pullulans MR의 풀루란 생산에 있어 세가지 변수 중 특히 초기 pH가 풀루란 생산에 가장 큰 영향을 미친다는 것을 확인하였다. 결과적으로 A. pullulans MR을 26℃에서 5일간 배양할 경우 반응표면분석법을 이용하여 얻은 최적 배양 조건은 sucrose 15%, soy peptone 0.4%, 초기 pH 7이였으며, 위의 배양 조건에서 예측할 수 있는 풀루란의 최대생산량은 47.6 g/L이었다. 이후 확인 실험을 위해 A. pullulans MR을 동일 조건으로 배양한 뒤 풀루란의 생산량을 관찰한 결과 48.9±3.4 g/L로 예측된 값과 근접한 값을 얻을 수 있었다. 추가적으로, sucrose의 경우 기존의 선행연구들에서도 A. pullulans의 풀루란 생산을 위한 탄소원으로 보고가 되었지만, 배지내 sucrose의 함량이 5%를 초과하게 될 경우 오히려 풀루란의 생성을 저해한다는 결과가 보고된 바가 있다(Shin 등, 1987b). 이는 배지내 sucrose의 함량이 높아 질수록 sucrose의 농도에 의한 삼투압 현상과 낮은 수분활성도로 인해 발생하며(Singh 등, 2009b), sucrose뿐만 아니라 glucose와 maltose에서도 같은 결과가 나타내는 것이 보고된 바 있다(Shin 등, 1987a). 하지만 이러한 보고들과 다르게 본 연구에서는 오히려 sucrose의 함량을 5%에서 15%까지 높아졌음에도 불구하고 풀루란의 생산이 오히려 증가하는 반대의 결과가 나타났다. 또한 A. pullulans MR의 풀루란 생산에 있어 가장 큰 영향을 미친 초기 배지의 pH의 경우 A. pullulans 균주에 따라 최적 pH가 pH 3.8부터 pH 7.5까지 다양하게 나타나는 것을 알 수 있었는데(Singh 등, 2018a; Wang 등, 2013; Shin 등, 1991) A. pullulans MR의 경우 비교적 중성인 pH 7에서 가장 많은 양의 풀루란이 생산된 것을 알 수 있었다. 본 실험을 통해 A. pullulans MR의 풀루란 대량생산 가능성을 확인하였고, 이를 활용하여 식품, 화장품, 제약 등 여러 광범위한 분야에서 미생물 유래 다당류인 풀루란의 산업적 활용이 가능할 것이라 생각한다.

Keywords

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