Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Enzymatic characterization of Paenibacillus amylolyticus xylanases GH10 and GH30 for xylan hydrolysis

Paenibacillus amylolyticus 유래 xylanase GH10 및 GH30의 xylan 가수분해 특성

  • Nam, Gyeong-Hwa (Division of Animal, Horticultural and Food Sciences, Graduate School of Chungbuk National University) ;
  • Jang, Myoung-Uoon (Division of Animal, Horticultural and Food Sciences, Graduate School of Chungbuk National University) ;
  • Kim, Min-Jeong (Division of Animal, Horticultural and Food Sciences, Graduate School of Chungbuk National University) ;
  • Lee, Jung-Min (Division of Animal, Horticultural and Food Sciences, Graduate School of Chungbuk National University) ;
  • Lee, Min-Jae (Division of Animal, Horticultural and Food Sciences, Graduate School of Chungbuk National University) ;
  • Kim, Tae-Jip (Division of Animal, Horticultural and Food Sciences, Graduate School of Chungbuk National University)
  • 남경화 (충북대학교 대학원 축산.원예.식품공학부 식품공학전공) ;
  • 장명운 (충북대학교 대학원 축산.원예.식품공학부 식품공학전공) ;
  • 김민정 (충북대학교 대학원 축산.원예.식품공학부 식품공학전공) ;
  • 이정민 (충북대학교 대학원 축산.원예.식품공학부 식품공학전공) ;
  • 이민재 (충북대학교 대학원 축산.원예.식품공학부 식품공학전공) ;
  • 김태집 (충북대학교 대학원 축산.원예.식품공학부 식품공학전공)
  • Received : 2016.11.29
  • Accepted : 2016.12.19
  • Published : 2016.12.31
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Abstract

The enzymatic degradation of xylans is the most versatile way to obtain the high value-added functional compounds or the fermentable sugars for renewable energy. The endo-${\beta}$-xylanases are the major enzymes which hydrolyze the internal ${\beta}$-1,4-linkages of xylan backbones to produce the mixtures of xylooligosaccharides including xylobiose and xylotriose. Among them, glucuronoxylanase GH30 can exclusively hydrolyze the internal ${\beta}$-1,4-linkages of xylans decorated with methylglucuronic acid branches. In the present study, two xylanolytic enzyme (PaXN_10 and PaGuXN_30) genes were cloned from Paenibacillus amylolyticus KCTC 3005, and expressed in Escherichia coli, respectively. PaXN_10 (38.7 kDa) belongs to the endo-${\beta}$-xylanases GH10 family, while PaGuXN_30 (58.5 kDa) is a member of glucuronoxylanase GH30. They share the same optimal reaction conditions at $50^{\circ}C$ and pH 7.0. Enzymatic characterization proposed that P. amylolyticus can utilize the hardwood glucuronoarabinoxylans via the cooperative actions of xylanases GH10 and GH30. The extracellular PaGuXN_30 is secreted into the medium and hydrolyzes glucuronoarabinoxylans to release a series of aldouronic acid mixtures with a methylglucuronic acid branch. The resultant products being transported into the microbial cell are successively degraded into the smaller xylooligosaccharides by the intracellular PaXN_10, which will be utilized for the cellular metabolism.

Xylan의 효소적 가수분해는 고부가가치 기능성 물질 또는 바이오에너지 생산을 위한 발효성 당을 얻는 가장 유용한 방법 중 하나이다. endo-${\beta}$-Xylanase는 xylan 주사슬 내부의 ${\beta}$-1,4-결합을 가수분해하여 xylobiose, xylotriose를 포함한 다양한 XOS를 생산하는 핵심 효소이다. 이들 효소 중에서 glucuronoxylanase GH30은 methylglucuronic acid가 측쇄에 수식된 xylan에 특이적으로 작용한다. 본 연구에서는 Paenibacillus amylolyticus KCTC 3005에서 유래한 2종의 xylan 가수분해효소(PaXN_10과 PaGuXN_30) 유전자를 클로닝하고, Escherichia coli에서 각각 발현시켰다. PaXN_10 (38.7 kDa)은 ${\beta}$-xylanase GH10 계열, PaGuXN_30 (58.5 kDa)은 glucuronoxylanase GH30에 해당하는 효소이며, $50^{\circ}C$와 pH 7.0에서 최대 활성을 나타내었다. 가수분해 특성 연구를 통해 P. amylolyticus가 목질계 glucuronoxylan을 분해하는 효소 시스템을 제안하였다. 세포 외로 분비되는 PaGuXN_30은 glucuroxylan을 가수분해하여 methylglucuronic acid 측쇄를 가지는 다양한 aldouronic acid mixtures를 생성하며, 이러한 분해산물은 세포 내로 이동하여 PaXN_GH10에 의해 xylose, xylobiose와 같은 저분자 XOS로 분해되어 세포 내 대사경로에 이용될 수 있다. 또한 이들 효소의 가수분해특성을 이용하여 다양한 탄수화물 소재 생산이 가능할 것으로 기대한다.

Keywords

References

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