Study on the Effects of Extraction and Purification Conditions on the Activity of Peroxidase and Polyphenol Oxidase from Chestnut Kernel and on Their Substrates

Gao Xiang; Han Mengmeng; Cai Ziyi; Zhang Xingyuan; Wang Fang; Pang Meixia; Qi Jinghua

doi:doi:10.11648/j.bio.20190702.13

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Study on the Effects of Extraction and Purification Conditions on the Activity of Peroxidase and Polyphenol Oxidase from Chestnut Kernel and on Their Substrates

Gao Xiang, Han Mengmeng, Cai Ziyi, Zhang Xingyuan, Wang Fang, Pang Meixia, Qi Jinghua

Received: 15 April 2019 Accepted: Published: 15 June 2019

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Abstract

Chestnut kernel is a woody grain with high nutritional value and utilized value, but it is susceptible to Browning. Previous studies have shown that phenol is the substrate of enzymes that lead to browning. In order to investigate the substrates that that leads to browning of peroxidase (POD) and polyphenol oxidase (PPO) of chestnut kernel. The extraction and purification methods and the main substrates of POD and PPO from chestnut kernel were investigated. First, the effects on enzyme activity of different kinds of buffer, temperature, time and pH value were studied; second, the salting out purification of different concentrations and the main substrates of POD and PPO from chestnut were studied. The results were shown that POD enzyme in chestnut could be obtained by extracting with sodium dihydrogen phosphate-dibasic sodium phosphate buffer (pH 6.8) at 4°C for 5 h. PPO enzyme in chestnut could be obtained by extracting with disodium hydrogen phosphate-citric acid buffer (pH 5.4) at 4°C for 5 h. The two enzymes could be purified best by 80% ammonium sulfate, and the activity of POD and PPO enzyme had been enhanced for 60% and 190% respectively. Gallic acid might be the main substrate of POD and PPO in browning of chestnut kernel.

DOI	10.11648/j.bio.20190702.13
Published in	American Journal of Bioscience and Bioengineering (Volume 7, Issue 2, April 2019)
Page(s)	40-45
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

Chestnut Kernel, Enzymatic Browning, Extraction, Purification, Substrate

References

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[4]	Geng Jiannuan. Analyse of Browning Factors during the Process of Chinese Chestnut [J]. Hubei Agricultural Sciences, 2014, 53 (08): 1879-1881.
[5]	ZENG Ting-ting, ZHANG Li-yan, RUI Han-ming. Study on enzymatic browning and blanching methods of chestnut [J]. Science and Technology of Food Industry, 2011, 32 (10): 110-113.
[6]	Chen Yalan, Wang Qing, Zhang Jian. Physical Activity of Transgenic Homozygous Triploid of Potatoes [J]. Chinese Potato Journal 2005, (1) 2000, (1): 24~25.
[7]	Sun Jian. Studies on the major substrates for polyphenol oxidases in pericarp tissues of litchi fruit [D]. Graduate University of Chinese Academy of Sciences PhD thesis, 2008.
[8]	Wang Jing, Qing Jinhua, Wu Yihong, et al. Study on the Factors that Related to Enzymatic Browning from the Castanea mollissima Blume Kernel [J]. Chinese Agricultural Science Bulletin, 2010, 26 (20): 73-79.
[9]	Department of Physiology, Department of Biology, East China Normal University. Plant Physiology Experiment Guide [M]. People's Education Press. 1980: 143~144.
[10]	Tian S P, Xu Y, Jiang A L, et a1. Changes in enzymatic activity and quality attributes of 1ate-mature peaches in response to controlled atmosphere conductions [J]. Agric Science in China, 2002, 1: 207~212.
[11]	Anderson J V, Fuerst P E, Hurkman W J, et al. Biochemical and genetic characterization of wheat (Tritcum spp.) kernel polyphenol oxidases [J]. Journal of Cereal Science, 2006, 44 (3): 353-367.
[12]	Liu Xiaoyang, Li Ling, Gao Guizhen. Influence of Light Intensity on POD, PAL and PPO Activity of Pyrus bretschneideri cv. Dangshan Su Fruit in Its Growth Phase [J]. Acta Laser Biology Sinica, 2008, (3): 39~53.
[13]	Wu ZX. Studies on the Enzymes that Related to cell wall Metabolism and Pericarp Browning in Longan [A]. Chinese Society for Horticultural Science. The Collected Papers of 7th Youth Symposium on Chinese Society for Horticultural Science [C]. Chinese Society for Horticultural Science: Chinese Society for Horticultural Science, 2006: 5.
[14]	Mao Linchun. Technical problems and countermeasures of chestnut processing [J]. Food Science and Technology. 2000, (1): 24~25.
[15]	SHI C H, DAI Y, XU X L, et al. The purification of polyphenol oxidase from tobacco [J]. Protein Expression and Purification, 2002, 24: 51~55.

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Gao Xiang, Han Mengmeng, Cai Ziyi, Zhang Xingyuan, Wang Fang, et al. (2019). Study on the Effects of Extraction and Purification Conditions on the Activity of Peroxidase and Polyphenol Oxidase from Chestnut Kernel and on Their Substrates. American Journal of Bioscience and Bioengineering, 7(2), 40-45. https://doi.org/10.11648/j.bio.20190702.13

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ACS Style

Gao Xiang; Han Mengmeng; Cai Ziyi; Zhang Xingyuan; Wang Fang, et al. Study on the Effects of Extraction and Purification Conditions on the Activity of Peroxidase and Polyphenol Oxidase from Chestnut Kernel and on Their Substrates. Am. J. BioSci. Bioeng. 2019, 7(2), 40-45. doi: 10.11648/j.bio.20190702.13

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AMA Style

Gao Xiang, Han Mengmeng, Cai Ziyi, Zhang Xingyuan, Wang Fang, et al. Study on the Effects of Extraction and Purification Conditions on the Activity of Peroxidase and Polyphenol Oxidase from Chestnut Kernel and on Their Substrates. Am J BioSci Bioeng. 2019;7(2):40-45. doi: 10.11648/j.bio.20190702.13

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@article{10.11648/j.bio.20190702.13,
  author = {Gao Xiang and Han Mengmeng and Cai Ziyi and Zhang Xingyuan and Wang Fang and Pang Meixia and Qi Jinghua},
  title = {Study on the Effects of Extraction and Purification Conditions on the Activity of Peroxidase and Polyphenol Oxidase from Chestnut Kernel and on Their Substrates},
  journal = {American Journal of Bioscience and Bioengineering},
  volume = {7},
  number = {2},
  pages = {40-45},
  doi = {10.11648/j.bio.20190702.13},
  url = {https://doi.org/10.11648/j.bio.20190702.13},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.bio.20190702.13},
  abstract = {Chestnut kernel is a woody grain with high nutritional value and utilized value, but it is susceptible to Browning. Previous studies have shown that phenol is the substrate of enzymes that lead to browning. In order to investigate the substrates that that leads to browning of peroxidase (POD) and polyphenol oxidase (PPO) of chestnut kernel. The extraction and purification methods and the main substrates of POD and PPO from chestnut kernel were investigated. First, the effects on enzyme activity of different kinds of buffer, temperature, time and pH value were studied; second, the salting out purification of different concentrations and the main substrates of POD and PPO from chestnut were studied. The results were shown that POD enzyme in chestnut could be obtained by extracting with sodium dihydrogen phosphate-dibasic sodium phosphate buffer (pH 6.8) at 4°C for 5 h. PPO enzyme in chestnut could be obtained by extracting with disodium hydrogen phosphate-citric acid buffer (pH 5.4) at 4°C for 5 h. The two enzymes could be purified best by 80% ammonium sulfate, and the activity of POD and PPO enzyme had been enhanced for 60% and 190% respectively. Gallic acid might be the main substrate of POD and PPO in browning of chestnut kernel.},
 year = {2019}
}

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TY  - JOUR
T1  - Study on the Effects of Extraction and Purification Conditions on the Activity of Peroxidase and Polyphenol Oxidase from Chestnut Kernel and on Their Substrates
AU  - Gao Xiang
AU  - Han Mengmeng
AU  - Cai Ziyi
AU  - Zhang Xingyuan
AU  - Wang Fang
AU  - Pang Meixia
AU  - Qi Jinghua
Y1  - 2019/06/15
PY  - 2019
N1  - https://doi.org/10.11648/j.bio.20190702.13
DO  - 10.11648/j.bio.20190702.13
T2  - American Journal of Bioscience and Bioengineering
JF  - American Journal of Bioscience and Bioengineering
JO  - American Journal of Bioscience and Bioengineering
SP  - 40
EP  - 45
PB  - Science Publishing Group
SN  - 2328-5893
UR  - https://doi.org/10.11648/j.bio.20190702.13
AB  - Chestnut kernel is a woody grain with high nutritional value and utilized value, but it is susceptible to Browning. Previous studies have shown that phenol is the substrate of enzymes that lead to browning. In order to investigate the substrates that that leads to browning of peroxidase (POD) and polyphenol oxidase (PPO) of chestnut kernel. The extraction and purification methods and the main substrates of POD and PPO from chestnut kernel were investigated. First, the effects on enzyme activity of different kinds of buffer, temperature, time and pH value were studied; second, the salting out purification of different concentrations and the main substrates of POD and PPO from chestnut were studied. The results were shown that POD enzyme in chestnut could be obtained by extracting with sodium dihydrogen phosphate-dibasic sodium phosphate buffer (pH 6.8) at 4°C for 5 h. PPO enzyme in chestnut could be obtained by extracting with disodium hydrogen phosphate-citric acid buffer (pH 5.4) at 4°C for 5 h. The two enzymes could be purified best by 80% ammonium sulfate, and the activity of POD and PPO enzyme had been enhanced for 60% and 190% respectively. Gallic acid might be the main substrate of POD and PPO in browning of chestnut kernel.
VL  - 7
IS  - 2
ER  -

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Author Information

Gao Xiang

Beijing Key Laboratory of Agricultural Product Detection and Control of Spoil age Organisms and Pesticide Residue, Beijing University of Agriculture, Beijing, China; The Teaching Group of Food Chemistry, Faculty of Food Science and Engineering, Beijing University of Agriculture, Beijing, China
Han Mengmeng

Beijing Key Laboratory of Agricultural Product Detection and Control of Spoil age Organisms and Pesticide Residue, Beijing University of Agriculture, Beijing, China; The Teaching Group of Food Chemistry, Faculty of Food Science and Engineering, Beijing University of Agriculture, Beijing, China
Cai Ziyi

Beijing Key Laboratory of Agricultural Product Detection and Control of Spoil age Organisms and Pesticide Residue, Beijing University of Agriculture, Beijing, China; The Teaching Group of Food Chemistry, Faculty of Food Science and Engineering, Beijing University of Agriculture, Beijing, China
Zhang Xingyuan

Beijing Key Laboratory of Agricultural Product Detection and Control of Spoil age Organisms and Pesticide Residue, Beijing University of Agriculture, Beijing, China; The Teaching Group of Food Chemistry, Faculty of Food Science and Engineering, Beijing University of Agriculture, Beijing, China
Wang Fang

Beijing Key Laboratory of Agricultural Product Detection and Control of Spoil age Organisms and Pesticide Residue, Beijing University of Agriculture, Beijing, China; The Teaching Group of Food Chemistry, Faculty of Food Science and Engineering, Beijing University of Agriculture, Beijing, China; Beijing Innovation Consortium of Swine Research System, Beijing, China
Pang Meixia

Beijing Key Laboratory of Agricultural Product Detection and Control of Spoil age Organisms and Pesticide Residue, Beijing University of Agriculture, Beijing, China; The Teaching Group of Food Chemistry, Faculty of Food Science and Engineering, Beijing University of Agriculture, Beijing, China; Beijing Innovation Consortium of Swine Research System, Beijing, China
Qi Jinghua

Beijing Key Laboratory of Agricultural Product Detection and Control of Spoil age Organisms and Pesticide Residue, Beijing University of Agriculture, Beijing, China; The Teaching Group of Food Chemistry, Faculty of Food Science and Engineering, Beijing University of Agriculture, Beijing, China; Beijing Innovation Consortium of Swine Research System, Beijing, China

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APA Style

Gao Xiang, Han Mengmeng, Cai Ziyi, Zhang Xingyuan, Wang Fang, et al. (2019). Study on the Effects of Extraction and Purification Conditions on the Activity of Peroxidase and Polyphenol Oxidase from Chestnut Kernel and on Their Substrates. American Journal of Bioscience and Bioengineering, 7(2), 40-45. https://doi.org/10.11648/j.bio.20190702.13

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ACS Style

Gao Xiang; Han Mengmeng; Cai Ziyi; Zhang Xingyuan; Wang Fang, et al. Study on the Effects of Extraction and Purification Conditions on the Activity of Peroxidase and Polyphenol Oxidase from Chestnut Kernel and on Their Substrates. Am. J. BioSci. Bioeng. 2019, 7(2), 40-45. doi: 10.11648/j.bio.20190702.13

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AMA Style

Gao Xiang, Han Mengmeng, Cai Ziyi, Zhang Xingyuan, Wang Fang, et al. Study on the Effects of Extraction and Purification Conditions on the Activity of Peroxidase and Polyphenol Oxidase from Chestnut Kernel and on Their Substrates. Am J BioSci Bioeng. 2019;7(2):40-45. doi: 10.11648/j.bio.20190702.13

Copy | Download

@article{10.11648/j.bio.20190702.13,
  author = {Gao Xiang and Han Mengmeng and Cai Ziyi and Zhang Xingyuan and Wang Fang and Pang Meixia and Qi Jinghua},
  title = {Study on the Effects of Extraction and Purification Conditions on the Activity of Peroxidase and Polyphenol Oxidase from Chestnut Kernel and on Their Substrates},
  journal = {American Journal of Bioscience and Bioengineering},
  volume = {7},
  number = {2},
  pages = {40-45},
  doi = {10.11648/j.bio.20190702.13},
  url = {https://doi.org/10.11648/j.bio.20190702.13},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.bio.20190702.13},
  abstract = {Chestnut kernel is a woody grain with high nutritional value and utilized value, but it is susceptible to Browning. Previous studies have shown that phenol is the substrate of enzymes that lead to browning. In order to investigate the substrates that that leads to browning of peroxidase (POD) and polyphenol oxidase (PPO) of chestnut kernel. The extraction and purification methods and the main substrates of POD and PPO from chestnut kernel were investigated. First, the effects on enzyme activity of different kinds of buffer, temperature, time and pH value were studied; second, the salting out purification of different concentrations and the main substrates of POD and PPO from chestnut were studied. The results were shown that POD enzyme in chestnut could be obtained by extracting with sodium dihydrogen phosphate-dibasic sodium phosphate buffer (pH 6.8) at 4°C for 5 h. PPO enzyme in chestnut could be obtained by extracting with disodium hydrogen phosphate-citric acid buffer (pH 5.4) at 4°C for 5 h. The two enzymes could be purified best by 80% ammonium sulfate, and the activity of POD and PPO enzyme had been enhanced for 60% and 190% respectively. Gallic acid might be the main substrate of POD and PPO in browning of chestnut kernel.},
 year = {2019}
}

Copy | Download

TY  - JOUR
T1  - Study on the Effects of Extraction and Purification Conditions on the Activity of Peroxidase and Polyphenol Oxidase from Chestnut Kernel and on Their Substrates
AU  - Gao Xiang
AU  - Han Mengmeng
AU  - Cai Ziyi
AU  - Zhang Xingyuan
AU  - Wang Fang
AU  - Pang Meixia
AU  - Qi Jinghua
Y1  - 2019/06/15
PY  - 2019
N1  - https://doi.org/10.11648/j.bio.20190702.13
DO  - 10.11648/j.bio.20190702.13
T2  - American Journal of Bioscience and Bioengineering
JF  - American Journal of Bioscience and Bioengineering
JO  - American Journal of Bioscience and Bioengineering
SP  - 40
EP  - 45
PB  - Science Publishing Group
SN  - 2328-5893
UR  - https://doi.org/10.11648/j.bio.20190702.13
AB  - Chestnut kernel is a woody grain with high nutritional value and utilized value, but it is susceptible to Browning. Previous studies have shown that phenol is the substrate of enzymes that lead to browning. In order to investigate the substrates that that leads to browning of peroxidase (POD) and polyphenol oxidase (PPO) of chestnut kernel. The extraction and purification methods and the main substrates of POD and PPO from chestnut kernel were investigated. First, the effects on enzyme activity of different kinds of buffer, temperature, time and pH value were studied; second, the salting out purification of different concentrations and the main substrates of POD and PPO from chestnut were studied. The results were shown that POD enzyme in chestnut could be obtained by extracting with sodium dihydrogen phosphate-dibasic sodium phosphate buffer (pH 6.8) at 4°C for 5 h. PPO enzyme in chestnut could be obtained by extracting with disodium hydrogen phosphate-citric acid buffer (pH 5.4) at 4°C for 5 h. The two enzymes could be purified best by 80% ammonium sulfate, and the activity of POD and PPO enzyme had been enhanced for 60% and 190% respectively. Gallic acid might be the main substrate of POD and PPO in browning of chestnut kernel.
VL  - 7
IS  - 2
ER  -

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