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An Acid-Adapted Endo-α-1,5-l-arabinanase for Pectin Releasing

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Abstract

An arabinanase gene was cloned by overlap-PCR from Penicillium sp. Y702 and expressed in Pichia pastoris. The recombinant enzyme was named AbnC702 with 20 U/mg of endo-arabinanase activity toward linear α-1,5-l-arabinan. The optimal pH and temperature of AbnC702 were 5.0 and 50 °C, respectively. The recombinant AbnC702 was highly stable at pH 5.0–7.0 and 50 °C. It could retain about 72.3 % of maximum specific activity at pH 5.0 after incubation for 2.5 h, which indicated AbnC702 was an acid-adapted enzyme. The K m and V max values were 24.8 ± 4.7 mg/ml and 88.5 ± 5.6 U/mg, respectively. A three-dimensional structure of AbnC702 was made by homology modeling, and the counting of acidic/basic amino residues within the region of 10 Å around the active site, as well the hydrogen bonds within the area of 5 Å around the active site, might theoretically interpret the acid adaptability of AbnC702. Analysis of hydrolysis products by thin layer chromatography (TLC) combined with high-performance liquid chromatography (HPLC) verified that the recombinant AbnC702 was an endo-1,5-α-l-arabinanase, which yielded arabinobiose and arabinotriose as major products. AbnC702 was applied in pectin extraction from apple pomace with synergistic action of α-L-arabinofuranosidase.

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Acknowledgments

This work was supported by National Special Fund for State Key Laboratory of Bioreactor Engineering (2060204) and partially supported by the National Natural Science Foundation of China (Nos. 21506057 and 21506057), the National High Technology Research and Development Program of China (No. 2013AA102109), and the Natural Science Foundation of Shanghai (No. 2013ZR1412100).

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Authors and Affiliations

  1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, People’s Republic of China

    Chong Lang, Rujian Yang, Ying Yang, Bei Gao, Li Zhao, Wei Wei, Hualei Wang, Jingli Xie & Dongzhi Wei

  2. Department of Food Science and Technology, School of Biotechnology, East China University of Science and Technology, Shanghai, 200237, People’s Republic of China

    Chong Lang, Rujian Yang, Ying Yang, Bei Gao, Li Zhao, Wei Wei, Hualei Wang, Jingli Xie & Dongzhi Wei

  3. Department of Food Science and Technology, Tokyo University of Marine Science and Technology, Tokyo, 108-8477, Japan

    Shingo Matsukawa

  4. Shanghai Collaborative Innovation Center for Biomanufacturing (SCICB), Shanghai, 200237, People’s Republic of China

    Jingli Xie & Dongzhi Wei

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  1. Chong Lang
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Correspondence to Jingli Xie.

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Lang, C., Yang, R., Yang, Y. et al. An Acid-Adapted Endo-α-1,5-l-arabinanase for Pectin Releasing. Appl Biochem Biotechnol 180, 900–916 (2016). https://doi.org/10.1007/s12010-016-2141-5

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