Xinjiang Agricultural Sciences ›› 2023, Vol. 60 ›› Issue (2): 432-439.DOI: 10.6048/j.issn.1001-4330.2023.02.021
• Horticultural Special Local Products·Plant Protection·Microbes·Soil Fertilizer· Water Saving Irrigation • Previous Articles Next Articles
Nurziya Yarmamat(), HAO Jingzhe(), JIA Wenjie, CHEN Haoyu, LUO Ying, JIA Peisong()
Received:
2022-05-27
Online:
2023-02-20
Published:
2023-03-31
Correspondence author:
HAO Jingzhe (1970-), researcher, research area: micropathology,(E-mail) urmqhjz@qq.com; JIA Peisong (1984-), associate researcher, research area: edible fungi research, (E-mail)jps-fly@163.com
Supported by:
努尔孜亚·亚力买买提(), 郝敬喆(), 贾文捷, 陈浩宇, 罗影, 贾培松()
通讯作者:
郝敬喆(1969-),男,江苏人,研究员,研究方向为微生物病理学,(E-mail)urmqhjz@qq.com; 贾培松(1984-),男,河北人,副研究员,研究方向为食用菌驯化与栽培,(E-mail)jps-fly@163.com
作者简介:
努尔孜亚·亚力买买提(1977-),女,新疆伊犁人,副研究员,研究方向为食用菌驯化与栽培, (E-mail)nrzy729@163.com
基金资助:
CLC Number:
Nurziya Yarmamat, HAO Jingzhe, JIA Wenjie, CHEN Haoyu, LUO Ying, JIA Peisong. Purification and Physicochemical Properties of Laccase from Fruiting Bodies of Agaricus balchaschensis[J]. Xinjiang Agricultural Sciences, 2023, 60(2): 432-439.
努尔孜亚·亚力买买提, 郝敬喆, 贾文捷, 陈浩宇, 罗影, 贾培松. 巴尔喀什蘑菇子实体漆酶纯化与理化特性及对酚类化合物的降解能力[J]. 新疆农业科学, 2023, 60(2): 432-439.
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URL: http://www.xjnykx.com/EN/10.6048/j.issn.1001-4330.2023.02.021
Fig.1 Ecological environment and development stage of A. balchaschensis Note: a. A. balchaschensis lake side habitat;b.The fruiting body 30 cm underground;c.Fruiting body shape;d.Spore print e:spore and Spore stalk(10×40X);f.Mycelial morphology
分离方式 Separation mode | 收集组分 Collect components | 收集量 Collection volume(mg) | 总活性 Total activity(U) | 比活力 Specific activity (U/mg) | 纯化倍数 Purification factor |
---|---|---|---|---|---|
DEAE-cellulose | Crude extract | 4142.75 | 198.85 | 0.048 | 1 |
CM-cellulose cation exchange | C1 | 19.89 | 1.01 | 0.051 | 1.06 |
C2 | 125.86 | 9.82 | 0.078 | 1.63 | |
C3 | 363.31 | 68.30 | 0.188 | 3.92 | |
Q-Sepharose anion exchange | Q1 | 83.82 | 31.26 | 0.373 | 7.77 |
Superdex 75 | SU1 | 6.4 | 108.93 | 17.02 | 354.58 |
Table 1 Results of laccase purification from A. balchaschensis mushroom
分离方式 Separation mode | 收集组分 Collect components | 收集量 Collection volume(mg) | 总活性 Total activity(U) | 比活力 Specific activity (U/mg) | 纯化倍数 Purification factor |
---|---|---|---|---|---|
DEAE-cellulose | Crude extract | 4142.75 | 198.85 | 0.048 | 1 |
CM-cellulose cation exchange | C1 | 19.89 | 1.01 | 0.051 | 1.06 |
C2 | 125.86 | 9.82 | 0.078 | 1.63 | |
C3 | 363.31 | 68.30 | 0.188 | 3.92 | |
Q-Sepharose anion exchange | Q1 | 83.82 | 31.26 | 0.373 | 7.77 |
Superdex 75 | SU1 | 6.4 | 108.93 | 17.02 | 354.58 |
Mushroom laccase | N-terminal sequence |
---|---|
BHBClaccase | SGGPEQNTTA |
Agaricus bisporus laccaseI | KTRTFDFDL |
Agaricus bisporus laccaseII | DTKTFNFDL |
Basidiomycete PM1laccase | SIGPVADLT |
Ceriporiopsis subvermispora laccaseI | AIGPVTDLE |
Lepista nuda laccase | AIGPAADLHIVNKDISPDGF |
Pleurotus ostreatus laccase | SIGPRGTLN |
Pleurotus eryngii laccaseI | AXKKLDFHI |
Pleurotus eryngii laccaseII | ATKKLDFHI |
Phlebia radiata laccase | SIGPVTDFH |
Pycnoporus cinnabarius laccase | AIGPVADLT |
Russula virescens laccase | AIGPTAELVV |
Trametes sp F1635 laccase | AIGPVADLTIINNAV |
Trametes versicolor laccaseI | AIGPVASLV |
Trametes versicolor laccaseII | GIGPVADLT |
Trametes versicolor laccaseIII | GIGPVADLT |
Tricholoma matsutake laccase | ADPVGIVGD |
Tricholoma giganteum laccase | DDPQQAVIDD |
Tricholoma mongolicum laccase | GIGPVADLYVGNRIL |
Table 2 N-terminal sequence comparison of Agaricus balchaschensis laccase and other mushroom laccases
Mushroom laccase | N-terminal sequence |
---|---|
BHBClaccase | SGGPEQNTTA |
Agaricus bisporus laccaseI | KTRTFDFDL |
Agaricus bisporus laccaseII | DTKTFNFDL |
Basidiomycete PM1laccase | SIGPVADLT |
Ceriporiopsis subvermispora laccaseI | AIGPVTDLE |
Lepista nuda laccase | AIGPAADLHIVNKDISPDGF |
Pleurotus ostreatus laccase | SIGPRGTLN |
Pleurotus eryngii laccaseI | AXKKLDFHI |
Pleurotus eryngii laccaseII | ATKKLDFHI |
Phlebia radiata laccase | SIGPVTDFH |
Pycnoporus cinnabarius laccase | AIGPVADLT |
Russula virescens laccase | AIGPTAELVV |
Trametes sp F1635 laccase | AIGPVADLTIINNAV |
Trametes versicolor laccaseI | AIGPVASLV |
Trametes versicolor laccaseII | GIGPVADLT |
Trametes versicolor laccaseIII | GIGPVADLT |
Tricholoma matsutake laccase | ADPVGIVGD |
Tricholoma giganteum laccase | DDPQQAVIDD |
Tricholoma mongolicum laccase | GIGPVADLYVGNRIL |
阳离子盐 Cationic salt | 相对酶活Relative enzyme activity(%) | ||
---|---|---|---|
5 mmol/L | 2.5 mmol/L | 1.25 mmol/L | |
Li+ | 62.12± 0.55 | 97.97 ± 0.41 | 98.42± 0.11 |
Na+ | 63.38 ± 0.07 | 97.75 ± 0.49 | 98.33 ± 0.1 |
Mg2+ | 60.63 ± 0.52 | 98.74 ± 0.49 | 97.21 ± 0.01 |
Al3+ | 58.96 ± 0.16 | 96.51 ± 0.77 | 97.57 ± 0.25 |
K+ | 50.52 ± 0.07 | 89.68 ± 0.67 | 91.37 ± 0.17 |
Ca2+ | 58.96 ± 0.78 | 96.71± 0.79 | 97.72± 0.13 |
Cr2+ | 62.22 ± 0.29 | 98.27 ± 0.08 | 99.53 ± 0.07 |
Mn2+ | 47.39 ± 0.42 | 92.54± 0.01 | 99.41 ± 0.18 |
Fe3+ | 63.58 ± 0.15 | 95.95 ± 0.23 | 97.82± 0.11 |
Cu2+ | 105.35 ± 0.15 | 101.77 ± 2.61 | 98.88 ± 2.07 |
Zn2+ | 60.11± 0.27 | 84.98 ± 0.41 | 98.72 ± 0.07 |
Cd2+ | 71.71 ± 0.17 | 76.42 ± 0.85 | 96.78 ± 0.01 |
Ba2+ | 61.28 ± 0.47 | 93.58 ± 0.01 | 99.41 ± 1.2 |
Hg2+ | 6.94 ± 0.05 | 9.55 ± 0.04 | 12.21 ± 0.84 |
Pb2+ | 85.32 ± 0.64 | 95.7 ± 0.69 | 94.66 ± 0.25 |
Table 3 Effect of metal ions rate of laccase from A. balchaschensis (%)
阳离子盐 Cationic salt | 相对酶活Relative enzyme activity(%) | ||
---|---|---|---|
5 mmol/L | 2.5 mmol/L | 1.25 mmol/L | |
Li+ | 62.12± 0.55 | 97.97 ± 0.41 | 98.42± 0.11 |
Na+ | 63.38 ± 0.07 | 97.75 ± 0.49 | 98.33 ± 0.1 |
Mg2+ | 60.63 ± 0.52 | 98.74 ± 0.49 | 97.21 ± 0.01 |
Al3+ | 58.96 ± 0.16 | 96.51 ± 0.77 | 97.57 ± 0.25 |
K+ | 50.52 ± 0.07 | 89.68 ± 0.67 | 91.37 ± 0.17 |
Ca2+ | 58.96 ± 0.78 | 96.71± 0.79 | 97.72± 0.13 |
Cr2+ | 62.22 ± 0.29 | 98.27 ± 0.08 | 99.53 ± 0.07 |
Mn2+ | 47.39 ± 0.42 | 92.54± 0.01 | 99.41 ± 0.18 |
Fe3+ | 63.58 ± 0.15 | 95.95 ± 0.23 | 97.82± 0.11 |
Cu2+ | 105.35 ± 0.15 | 101.77 ± 2.61 | 98.88 ± 2.07 |
Zn2+ | 60.11± 0.27 | 84.98 ± 0.41 | 98.72 ± 0.07 |
Cd2+ | 71.71 ± 0.17 | 76.42 ± 0.85 | 96.78 ± 0.01 |
Ba2+ | 61.28 ± 0.47 | 93.58 ± 0.01 | 99.41 ± 1.2 |
Hg2+ | 6.94 ± 0.05 | 9.55 ± 0.04 | 12.21 ± 0.84 |
Pb2+ | 85.32 ± 0.64 | 95.7 ± 0.69 | 94.66 ± 0.25 |
底物 Substrate | 波长 Wavel ength | 相对酶活 Relative enzyme activity |
---|---|---|
咖啡酸 Caffeic acid | 420 | 0.930±0.03 |
没食子酸 Gallic acid | 450 | 0.60±0.04 |
甲苯胺 Toluidine | 366 | 15.77±0.01 |
联苯胺 Benzidine | 366 | 8.10±0.03 |
邻苯二酚 Catechol | 392 | 1.00±0.06 |
2,6二甲氧基酚 2,6 dimethoxyhenool | 460 | 1.10±0.05 |
愈创木酚 Guaiacol | 470 | 0.90±0.03 |
对苯二酚 Hydroquinone | 248 | 34±0.01 |
ABTS | 405 | 100±0.25 |
Table 4 Substrate diversity of laccase from A. balchaschensis mushroom
底物 Substrate | 波长 Wavel ength | 相对酶活 Relative enzyme activity |
---|---|---|
咖啡酸 Caffeic acid | 420 | 0.930±0.03 |
没食子酸 Gallic acid | 450 | 0.60±0.04 |
甲苯胺 Toluidine | 366 | 15.77±0.01 |
联苯胺 Benzidine | 366 | 8.10±0.03 |
邻苯二酚 Catechol | 392 | 1.00±0.06 |
2,6二甲氧基酚 2,6 dimethoxyhenool | 460 | 1.10±0.05 |
愈创木酚 Guaiacol | 470 | 0.90±0.03 |
对苯二酚 Hydroquinone | 248 | 34±0.01 |
ABTS | 405 | 100±0.25 |
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