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Soil Enzymes and Their Research Methods

  • Structural Biochemistry

  • https://enzyme.expasy.org/

  • https://www.brenda-enzymes.org
  • https://www.ebi.ac.uk/thornton-srv/databases/enzymes/
  • https://www.cazy.org/
  • https://doi.org/10.1093/nar/gkae1045 - CAZac
  • https://doi.org/10.1007/978-3-642-46687-8
  • https://doi.org/10.2136/sssabookser9
  • https://doi.org/10.1016/B978-0-12-513840-6.X5014-9
  • https://www.iso.org/standard/67074.html
  • https://www.iso.org/standard/74739.html

Experiment

Litter Enzyme

  • In-situ sampling; Season

    https://doi.org/10.1016/S0038-0717(02)00074-3
https://doi.org/10.1016/j.soilbio.2019.107690
https://doi.org/10.1016/j.soilbio.2009.11.005
https://doi.org/10.1016/j.soilbio.2004.07.018

  • In-situ incubated litterbag

    https://doi.org/10.1016/j.scitotenv.2024.171078
https://doi.org/10.1016/j.soilbio.2019.03.016
https://doi.org/10.1016/j.soilbio.2021.108230
https://doi.org/10.1016/j.scitotenv.2022.155743

  • Litter controlled incubation

    https://doi.org/10.1016/j.pedobi.2016.09.002
https://doi.org/10.1016/j.geoderma.2025.117283
https://doi.org/10.1016/j.soilbio.2011.12.009

Price of enzyme activity substrate

Enzyme

CAS Name Price Web
207738-08-7 TMB 169RMB/250mg https://www.beyotime.com/product/ST746.htm
30931-67-0 ABTS 176RMB/1g https://www.aladdin-e.com/zh_cn/a109612.html
3482-57-3 pNP-β-D-cellobioside 1145RMB/1g https://www.sunwaypharm.cn/product/4151.html
3459-18-5 pNP-N-acetyl-β-D-glucosaminide 372RMB/1g https://www.leyan.com/3459-18-5.html
645-15-8 bis-pNPP 34RMB/1g https://www.aladdin-e.com/zh_cn/b113674.html
4264-83-9 pNPP 30RMB/1g https://www.shyuanye.com/goods-S19138.html
31796-55-1 L-Alanine pNA hydrochloride 480RMB/1g https://www.shyuanye.com/goods-S45855.html
40127-11-5 L-Arginine pNA dihydrochloride 75RMB/1g https://www.shyuanye.com/goods-S46869.html
67953-08-6 L-Glutamic acid γ-(pNA) hydrochloride 635RMB/250mg https://www.macklin.cn/products/S931805
103213-34-9 glycine pNA 5915RMB/1g https://www.macklin.cn/products/P907100
4178-93-2 L-leucine pNA 218RMB/1g https://www.jkchemical.com/product/46810
40492-96-4 L-lysine pNA dihydrobromide 1124RMB/1g https://www.macklin.cn/products/L919901
6042-04-2 L-methionine pNA 616RMB/1g https://www.macklin.cn/products/S970788
108321-19-3 L-proline pNA trifluoroacetic acid salt 98RMB/1g https://www.aladdin-e.com/zh_cn/s138002.html

Microbial biomass

CAS Name Price Web
3810-74-0 Streptomycin sulfate 39RMB/10g https://www.macklin.cn/products/S6153
66-81-9 Cycloheximide 540RMB/1g https://shyuanye.com/goods-W13044.html

Soil Enzyme Activity

EC 1.11.1.7 - Broad-spectrum peroxidase - 广谱过氧化物酶

  • https://doi.org/10.1016/j.soilbio.2025.109913

EC 1.10.3.2 - Phenol oxidases - 漆酶（酚氧化酶）

  • https://doi.org/10.1016/j.mimet.2007.09.020
  • https://doi.org/10.1016/j.soilbio.2025.109861
  • https://doi.org/10.1016/j.chemosphere.2012.03.019
  • https://doi.org/10.1016/j.soilbio.2007.09.013
  • https://doi.org/10.1016/j.soilbio.2004.07.013
  • https://doi.org/10.1016/j.soilbio.2009.10.014

EC 1.11.1.13 - Manganese peroxidase - 锰过氧化物酶

  • https://doi.org/10.1016/j.soilbio.2012.07.007

EC 1.11.1.14 - Lignin peroxidase - 木质素过氧化物酶

  • https://doi.org/10.1016/j.soilbio.2012.07.007
  • https://doi.org/10.1016/j.soilbio.2009.10.014

EC 3.2.1.4 - Endoglucanases - 内切葡聚糖酶（羧甲基纤维素酶）

  • https://doi.org/10.1016/j.soilbio.2011.03.019
  • https://doi.org/10.1016/0038-0717(94)90216-X
  • https://doi.org/10.1016/j.soilbio.2008.01.015
  • https://doi.org/10.1016/j.heliyon.2024.e24022
  • https://doi.org/10.1016/0038-0717(92)90074-8
  • https://doi.org/10.1016/j.jgeb.2016.12.001

EC 3.2.1.91 - Exoglucanases - 外切葡聚糖酶（纤维二糖水解酶）

  • https://doi.org/10.1016/j.soilbio.2011.03.019
  • https://doi.org/10.1016/j.heliyon.2024.e24022
  • https://doi.org/10.1016/j.soilbio.2024.109416
  • https://doi.org/10.1016/j.soilbio.2004.09.005

EC 3.2.1.21 - β-glucosidases - β-葡萄糖苷酶

  • https://doi.org/10.1016/j.soilbio.2011.03.019
  • https://doi.org/10.1016/j.heliyon.2024.e24022
  • https://doi.org/10.1016/j.soilbio.2008.05.001
  • https://doi.org/10.1016/j.soilbio.2017.12.020
  • https://doi.org/10.1016/j.soilbio.2016.11.003

EC 3.2.1.8 - Xylanase - 木聚糖酶

  • https://doi.org/10.1016/S0038-0717(99)00092-9

EC 3.2.1.37 - β-D-xylosidase - 木糖苷酶

  • https://doi.org/10.1016/S0038-0717(99)00092-9

EC 3.2.1.55 - α-L-arabinosidase - α-L-阿拉伯糖苷酶

  • https://doi.org/10.1016/S0038-0717(99)00092-9

EC 3.2.1.31 - β-D-glucuronidase - β-D-葡糖醛酸苷酶

  • https://doi.org/10.1016/S0038-0717(99)00092-9

EC 3.1.1.6 - Acetyl xylan esterase - 乙酰木聚糖酯酶

  • https://doi.org/10.1016/S0038-0717(99)00092-9

EC 3.2.1.23 - β-Galactosidase - β-半乳糖苷酶

方法：鲜土经2mm筛分；使用去离子水作为溶剂，土壤（g）与水（mL）比例为4:25（凋落物与水比例为1:25），即在50ml尖底离心管内放入4g土壤和25mL去离子水（可使用30mL注射器）；使用涡旋振荡器至离心管内土壤分散，再至平板摇床以250rpm振荡10min，移液前再次涡旋振荡至形成土壤悬液（管底无颗粒沉淀）；96孔聚苯乙烯微孔板中，先使用50μL移液器将底物25μL移至微孔，再使用200μL移液器将125μL土壤悬液移至微孔，共计3个重复（添加底物）和1个空白（无底物，且此处不添加去离子水）；在37°C（35°C）孵育2h；孵育后在每个微孔加入25μL CaCl2和100μL pH12 100 mmol/L Tris溶液（枪头伸入微孔使溶液均质化）以停止反应，并在对照组添加底物25μL（添加底物需换枪头以防止污染）；将微孔板覆盖封板膜，以20°C 1500g离心5min，并将200μL上清液转移至新微孔板，于405nm比色获得吸光度。

终止溶液：pH12 100 mmol/L Tris溶液，将Tris（CAS 77-86-1 Mw 121.14）12.11g溶解于800mL去离子水中，并用5mol/L 氢氧化钠调节至pH12，定容至1000mL，在4°C储存不超过1month；0.5mol/L CaCl2溶液，将14.7g氯化钙二水合物（CAS 10035-04-8 Mw 147.01）溶解在200mL去离子水中，在4°C储存不超过1month；使用精度为0.001g的天平，下同。

底物溶液：0.05mol/L 对硝基苯基-β-D-吡喃半乳糖苷（CAS 3150–24–1 Mw 301,25）；将0.753g溶解于50mL 去离子水中。

标准溶液：3.6mmol/L 对硝基苯酚（PNP CAS 100-02-7）溶液：对硝基苯酚 Mw 139.11 10mg （100mg），去离子水 20mL（200mL）；使用时需稀释10倍，即工作浓度为0.36 mM。

校准曲线：需一式三份，通过使用移液器2-3次抽吸以均质液体；使用浓度为0.36 mmol/L 的PNP溶液，分别配置0 mmol/mL、14 mmol/mL、29 mmol/mL、72 mmol/mL、140 mmol/mL、220 mmol/mL、290 mmol/mL、360 mmol/mL的多孔板中分配所需体积，如表所示；之后加入25μL去离子水，25μL CaCl2溶液，100μL pH12 100mmol/L Tris溶液，均质化后转移200μl至新微孔板，在405nm读取吸光度。

PNP (nmol/ml) 0 14 29 72 140 220 290 360
PNP (μl) 0 5 10 25 50 75 100 125
Water (μl) 125 120 115 100 75 50 25 0
  • https://doi.org/10.3389/fpls.2018.00539
  • https://doi.org/10.1016/j.chemosphere.2009.03.014
  • https://doi.org/10.1016/j.soilbio.2010.06.018
  • https://doi.org/10.1104/pp.100.3.1318 - 参与果胶水解
  • https://doi.org/10.1093/jxb/erm037 - 作用于初生细胞壁，参与果胶水解

EC 3.2.1.1 - Amylose endo-a-1,4-glucanase - 内切-α-1，4-葡聚糖酶（α-淀粉酶）

  • https://doi.org/10.1016/S0038-0717(00)00142-5 - 凋落物酶活性

EC 3.2.1.26 - Fructofuranosidase - 呋喃果糖苷酶（蔗糖酶）

  • https://doi.org/10.1016/j.soilbio.2014.07.004

EC 3.1.1.2 - Arylesterase - 芳基酯酶

  • https://doi.org/10.1016/j.soilbio.2009.01.003

EC 3.2.1.30 - N-acetyl-β-D-glucosaminidase - N-乙酰-β-D-氨基葡萄糖苷酶

  • https://doi.org/10.1016/S0038-0717(00)00034-1
  • https://doi.org/10.1016/j.soilbio.2024.109416

EC - Tyrosinases - 酪氨酸酶

  • https://doi.org/10.1016/j.soilbio.2025.109903

EC 3.1.4.1 - Phosphodiesterase - 磷酸二酯酶

  • https://doi.org/10.1016/j.soilbio.2012.02.008

EC 3.1.3.(8,26) - Phytase - 植酸酶

  • https://doi.org/10.1016/j.soilbio.2009.12.002

EC 3.1.6.1 - Arylsulfatase - 芳基硫酸酯酶

  • https://doi.org/10.1016/j.pedsph.2022.06.006

EC 1.8.3.1 - Sulphite Xidase - 亚硫酸盐氧化酶

  • https://doi.org/10.1007/s42729-025-02392-1
  • https://doi.org/10.1016/j.pedsph.2022.06.006

Soil Microbial Biomass

Chloroform fumigation-extraction - 熏蒸浸提法

  • https://doi.org/10.1016/j.soilbio.2011.01.006

Ecoenzymatic Stoichiometry - 生态酶化学计量

  • https://doi.org/10.1016/j.geoderma.2022.116242
  • https://doi.org/10.1016/j.soilbio.2025.109886

Soil Enzyme Kinetics - 酶动力学

  • https://doi.org/10.1111/j.1365-2486.2011.02545.x
  • https://doi.org/10.1016/j.geodrs.2021.e00472
  • https://doi.org/10.1016/j.scitotenv.2019.136383
  • https://doi.org/10.1016/j.soilbio.2017.07.005
  • https://doi.org/10.1016/j.soilbio.2019.107656
  • https://doi.org/10.1007/s00374-012-0723-0
  • https://doi.org/10.1007/s10533-015-0086-3
  • https://doi.org/10.1016/j.soilbio.2012.01.011
  • https://doi.org/10.1016/j.soilbio.2018.07.021
  • https://doi.org/10.1111/j.1365-2486.2011.02615.x
  • https://doi.org/10.1002/jpln.202400242
  • https://doi.org/10.1111/gcb.12596
  • https://doi.org/10.1016/j.scitotenv.2023.163542
  • https://doi.org/10.1016/j.ejsobi.2018.11.004

Soil Enzyme Thermodynamic - 酶热力学

  • https://doi.org/10.1016/j.soilbio.2006.08.002

Factor

Tannin - 单宁

  • https://doi.org/10.1016/j.soilbio.2016.12.027

Sugar - 糖

  • https://doi.org/10.1016/j.soilbio.2015.07.021

Metal - 金属

  • https://doi.org/10.1016/j.soilbio.2008.09.011

    • 多价金属阳离子（Fe、Al）会抑制植酸酶（P）的活性
    • 酶底物与能够激活酶的二价阳离子（Mn、Co）、单价金属阳离子（Na、K）相关联

Understory - 林下植被

  • https://doi.org/10.1016/j.geoderma.2025.117672
  • https://doi.org/10.1016/j.actao.2024.104020