Appl Environ Microbiol. 1995 Oct;61(10):3687-94. doi: 10.1128/aem.61.10.3687-3694.1995.
Protoplasts of the monokaryotic strain 52J of Trametes versicolor were treated with UV light and screened for the inability to produce a colored precipitate on guaiacol-containing agar plates. Mutants unable to oxidize guaiacol had absent or very low secretion of laccase and manganese peroxidase (MnP) proteins. All isolates unable to secrete MnP were also unable to bleach or delignify kraft pulp. One mutant strain, M49, which grew normally but did not oxidize guaiacol, was tested further with a number of other substrates whose degradation has been associated with delignification by white rot fungi. Compared with the parent, 52J, mutant M49, secreting no MnP and low laccase, could not brighten or delignify kraft pulp, produced less ethylene from 2-keto methiolbutyric acid, released much less (sup14)CO(inf2) from [(sup14)C]DHP (a synthetic lignin-like polymerizate), and produced much less methanol from pulp. This mutant also displayed decreased abilities to oxidize the dyes poly B-411, poly R-478, and phenol red compared with the wild-type strain and was also unable to decolorize kraft bleachery effluent or mineralize its organochlorine. Addition of purified MnP in conjunction with H(inf2)O(inf2), MnSO(inf4), and an Mn(III) chelator to M49 cultures partially restored methanol production, pulp delignification, and biobleaching in some cases.
糙皮侧耳 52J 单核菌株的原生质体经紫外线处理后，在含有愈创木酚的琼脂平板上筛选不能产生有色沉淀的突变体。不能氧化愈创木酚的突变体，漆酶和锰过氧化物酶（MnP）蛋白的分泌缺失或非常低。所有不能分泌 MnP 的分离物也不能漂白或脱木质素硫酸盐浆。一株突变株 M49 可以正常生长但不能氧化愈创木酚，进一步用许多其他底物进行测试，这些底物的降解与白腐真菌的木质素脱除有关。与亲本 52J 相比，不分泌 MnP 和低漆酶的突变体 M49 不能使硫酸盐浆增白或脱木质素，从 2-酮基硫醇丁酸产生的乙烯较少，从 [(sup14)C]DHP（一种合成的木质素类似物聚合体）释放的 (sup14)CO(inf2) 较少，从纸浆中产生的甲醇较少。与野生型菌株相比，该突变体对多 B-411、多 R-478 和酚红等染料的氧化能力也降低，也不能使硫酸盐浆漂白白腐液脱色或矿化其有机氯。在 M49 培养物中添加纯化的 MnP 以及 H(inf2)O(inf2)、MnSO(inf4)和 Mn(III)螯合剂部分恢复了甲醇的产生、纸浆脱木质素和生物漂白作用。
