草业学报 ›› 2026, Vol. 35 ›› Issue (7): 228-239.DOI: 10.11686/cyxb2025257
• 综合评述 • 上一篇
收稿日期:2025-06-26
修回日期:2025-10-23
出版日期:2026-07-20
发布日期:2026-05-21
通讯作者:
周斐然
作者简介:Corresponding author. E-mail: 710298044@qq.com基金资助:
De-zhi LONG(
), Zeng-zhao LIU, Fei-ran ZHOU(
)
Received:2025-06-26
Revised:2025-10-23
Online:2026-07-20
Published:2026-05-21
Contact:
Fei-ran ZHOU
摘要:
中国作为棉花生产与消费大国,每年产生大量棉副产品,但其含有的游离棉酚等抗营养因子严重限制了其在饲料中的高值化应用。传统物理化学脱毒方法存在成本高、效率低及潜在污染等问题。漆酶作为一种多铜氧化酶,凭借其广谱底物特异性和高效氧化能力,能够通过催化棉酚分子内环化反应,将毒性醛基转化为稳定的呋喃结构,实现棉酚的绿色降解。系统综述了棉酚的化学特性及其对动物肝毒性、生殖毒性与氧化应激的影响,深入解析了漆酶的三维结构、铜簇催化机制及介体辅助氧化途径,总结了pH、温度、酶浓度等关键因素对降解效率的调控规律。结果发现,漆酶处理可显著缓解棉酚引起的肝损伤与生殖功能障碍,恢复抗氧化酶活性。此外,通过天然菌株筛选、基因工程改造及极端环境微生物开发,漆酶的生产效率与稳定性持续提升,其在棉粕脱毒中的应用案例已验证其工业化潜力。未来,漆酶技术有望推动棉副产品资源化利用,降低饲料成本,并衍生高附加值产物,为实现棉产业可持续发展提供关键技术支撑。
龙德智, 刘增照, 周斐然. 漆酶在棉副产品饲料开发中的研究进展[J]. 草业学报, 2026, 35(7): 228-239.
De-zhi LONG, Zeng-zhao LIU, Fei-ran ZHOU. Progress in research on the use of laccase to produce feed from cotton by-products[J]. Acta Prataculturae Sinica, 2026, 35(7): 228-239.
图2 基于底物漆酶的氧化循环[6]A: 直接底物氧化Direct oxidation of substrates; B: 化学催化底物氧化Chemically catalyzed substrate oxidation; C: 多酶偶联氧化Multienzymatic coupled oxidation. red(reduced)指失去电子过程,ox(oxidized)指得到电子过程。red (reduced) refers to the process of losing electrons; ox (oxidized) refers to the process of obtaining electrons.
Fig.2 Laccase-catalyzed redox cycles for substrate oxidation
| 酶源类型Enzyme source type | 代表酶Representative enzyme | pH值 pH value |
|---|---|---|
| 真菌漆酶Fungal laccase | 云芝漆酶Trametes versicolor laccase[ | 4.0~6.0 |
| 细菌漆酶Bacterial laccase | 植物乳杆菌漆酶Lactobacillus plantarum laccase[ | 4.0~7.0 |
| 微生物发酵Microbial fermentation | 芽孢杆菌漆酶Bacillus licheniformis laccase[ | 6.0~7.5 |
表1 不同来源漆酶催化pH
Table 1 pH catalyzed by laccases from different sources
| 酶源类型Enzyme source type | 代表酶Representative enzyme | pH值 pH value |
|---|---|---|
| 真菌漆酶Fungal laccase | 云芝漆酶Trametes versicolor laccase[ | 4.0~6.0 |
| 细菌漆酶Bacterial laccase | 植物乳杆菌漆酶Lactobacillus plantarum laccase[ | 4.0~7.0 |
| 微生物发酵Microbial fermentation | 芽孢杆菌漆酶Bacillus licheniformis laccase[ | 6.0~7.5 |
| 酶源类型Enzyme source type | 不同酶源代表物Representative enzymes from different sources | 适宜温度Suitable temperature |
|---|---|---|
| 真菌漆酶Fungal laccase | 变色栓菌同工酶T. versicolor isozymes[ | 60~70 ℃ |
| 杂色云芝同工酶Coriolus versicolor isozymes[ | 55~60 ℃ | |
| 细菌漆酶Bacteria laccase | 地衣芽孢杆菌B. licheniformis[ | 60 ℃左右About 60 ℃ |
| 龙舌兰芽孢杆菌B. tequilensis[ | 85 ℃左右About 85 ℃ | |
| 植物漆酶Plant laccase | 日本漆树Toxicodendron vernicifluum[ | 28 ℃左右About 28 ℃ |
| 基因工程漆酶Genetically engineered laccase | 重组漆酶Recombinant laccase[ | 55 ℃左右About 55 ℃ |
表2 不同来源漆酶催化适宜温度
Table 2 Suitable temperature for laccase catalysis from different sources
| 酶源类型Enzyme source type | 不同酶源代表物Representative enzymes from different sources | 适宜温度Suitable temperature |
|---|---|---|
| 真菌漆酶Fungal laccase | 变色栓菌同工酶T. versicolor isozymes[ | 60~70 ℃ |
| 杂色云芝同工酶Coriolus versicolor isozymes[ | 55~60 ℃ | |
| 细菌漆酶Bacteria laccase | 地衣芽孢杆菌B. licheniformis[ | 60 ℃左右About 60 ℃ |
| 龙舌兰芽孢杆菌B. tequilensis[ | 85 ℃左右About 85 ℃ | |
| 植物漆酶Plant laccase | 日本漆树Toxicodendron vernicifluum[ | 28 ℃左右About 28 ℃ |
| 基因工程漆酶Genetically engineered laccase | 重组漆酶Recombinant laccase[ | 55 ℃左右About 55 ℃ |
酶源 Enzyme source | 目标底物 Target substrate | 典型酶浓度 Typical enzyme concentration | 温度 Temperature (℃) | pH | 反应时间 Reaction time (h) | 棉酚降解率 Gossypol degradation rate (%) | 介体使用 Mediator usage |
|---|---|---|---|---|---|---|---|
| 解淀粉芽孢杆菌Bacillus amyloliquefaciens[ | 游离棉酚 Free gossypol | 101.56 U·mL-1 | 55 | 4 | 2 | 94.57 | - |
| 98.73 | 2,2'-联氮-双(3-乙基苯并噻唑啉-6-磺酸) 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) | ||||||
| 细菌漆酶Spore coat protein A laccase (CotA laccase)[ | 脱脂棉籽粕中游离棉酚Free gossypol of defatted cottonseed meal | - | 37 | 7 | 1 | 100.00 | - |
真菌漆酶 Fungal laccase | 多酚类化合物Polyphenolic compounds | 广泛 Extensive | 50~60 | 酸性 Acidic (4.0~6.5) | 几十分钟至数小时不等Vary from tens of minutes to several hours | 因酶和条件而异Depends on the enzyme and conditions | - |
商业漆酶 Patented laccase[ | 棉籽蛋白中游离棉酚Free gossypol of cottonseed protein | 1000~40000 U·L-1 | 20~40 | - | 10~30 | 未明确量化 Not clearly specified | 烟酰胺腺嘌呤二核苷酸(NAD)或过氧化氢(H2O2) Nicotinamide adenine dinucleotide or hydrogen peroxide |
表3 不同来源漆酶降解棉酚酶浓度与反应时间及条件
Table 3 Enzyme concentration, reaction time, and conditions for gossypol degradation by laccases from different sources
酶源 Enzyme source | 目标底物 Target substrate | 典型酶浓度 Typical enzyme concentration | 温度 Temperature (℃) | pH | 反应时间 Reaction time (h) | 棉酚降解率 Gossypol degradation rate (%) | 介体使用 Mediator usage |
|---|---|---|---|---|---|---|---|
| 解淀粉芽孢杆菌Bacillus amyloliquefaciens[ | 游离棉酚 Free gossypol | 101.56 U·mL-1 | 55 | 4 | 2 | 94.57 | - |
| 98.73 | 2,2'-联氮-双(3-乙基苯并噻唑啉-6-磺酸) 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) | ||||||
| 细菌漆酶Spore coat protein A laccase (CotA laccase)[ | 脱脂棉籽粕中游离棉酚Free gossypol of defatted cottonseed meal | - | 37 | 7 | 1 | 100.00 | - |
真菌漆酶 Fungal laccase | 多酚类化合物Polyphenolic compounds | 广泛 Extensive | 50~60 | 酸性 Acidic (4.0~6.5) | 几十分钟至数小时不等Vary from tens of minutes to several hours | 因酶和条件而异Depends on the enzyme and conditions | - |
商业漆酶 Patented laccase[ | 棉籽蛋白中游离棉酚Free gossypol of cottonseed protein | 1000~40000 U·L-1 | 20~40 | - | 10~30 | 未明确量化 Not clearly specified | 烟酰胺腺嘌呤二核苷酸(NAD)或过氧化氢(H2O2) Nicotinamide adenine dinucleotide or hydrogen peroxide |
酶源类型 Enzyme source type | 代表菌株 Representative strains | 产酶周期 Enzyme production cycle | 酶活性 Enzyme activity (U·mL-1) | 优势 Advantage | 局限性 Shortcoming |
|---|---|---|---|---|---|
| 天然真菌Natural fungal | 云芝T. versicolor[ | 7~14 d | 50~200 | 酶系丰富,底物广谱Rich enzyme system, broad substrate spectrum | 周期长,需要诱导剂Long cycle, need inducer |
| 基因工程细菌Genetically engineered bacterium | 枯草芽孢杆菌Bacillus subtilis[ | 24~48 h | 100~500 | 发酵快,耐高温/碱性Fast fermentation, high temperature/alkaline resistance | 需优化分泌表达Secretory expression needs to be optimized |
| 重组酵母Recombinant yeast | 毕赤酵母Pichia pastoris[ | 3~5 d | 200~800 | 糖基化修饰,稳定性高Glycosylation modification, high stability | 成本较高Expensive cost |
| 极端环境菌株Extreme environment strains | 嗜热芽孢杆菌Bacillus thermophilus[ | 48~72 h | 80~300 | 耐高温(>80 ℃),适合工业反应High temperature resistance (>80 ℃), suitable for industrial reactions | 底物特异性窄Narrow substrate specificity |
表4 不同酶源的性能比较
Table 4 Performance comparison of different enzyme sources
酶源类型 Enzyme source type | 代表菌株 Representative strains | 产酶周期 Enzyme production cycle | 酶活性 Enzyme activity (U·mL-1) | 优势 Advantage | 局限性 Shortcoming |
|---|---|---|---|---|---|
| 天然真菌Natural fungal | 云芝T. versicolor[ | 7~14 d | 50~200 | 酶系丰富,底物广谱Rich enzyme system, broad substrate spectrum | 周期长,需要诱导剂Long cycle, need inducer |
| 基因工程细菌Genetically engineered bacterium | 枯草芽孢杆菌Bacillus subtilis[ | 24~48 h | 100~500 | 发酵快,耐高温/碱性Fast fermentation, high temperature/alkaline resistance | 需优化分泌表达Secretory expression needs to be optimized |
| 重组酵母Recombinant yeast | 毕赤酵母Pichia pastoris[ | 3~5 d | 200~800 | 糖基化修饰,稳定性高Glycosylation modification, high stability | 成本较高Expensive cost |
| 极端环境菌株Extreme environment strains | 嗜热芽孢杆菌Bacillus thermophilus[ | 48~72 h | 80~300 | 耐高温(>80 ℃),适合工业反应High temperature resistance (>80 ℃), suitable for industrial reactions | 底物特异性窄Narrow substrate specificity |
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