Selective synthesis of the resveratrol analogue 4,4 '-dihydroxy-trans-stilbene and stilbenoids modification by fungal peroxygenases

Aranda Carmen; Ullrich Rene; Kiebist JanScheibner Katrindel Rio Jose C.Hofrichter MartinMartinez Angel T.Gutierrez Ana

摘要

This work gives first evidence that the unspecific peroxygenases (UPOs) from the basidiomycetes Agrocybe aegerita (AaeUPO), Coprinopsis cinerea (rCciUPO) and Marasmius rotula (MroUPO) are able to catalyze the regioselective hydroxylation of trans-stilbene to 4,4-dihydroxy-trans-stilbene (DHS), a resveratrol (RSV) analogue whose preventive effects on cancer invasion and metastasis have very recently been shown. Nearly complete transformation of substrate (yielding DHS) was achieved with the three enzymes tested, using H2O2 as the only co-substrate, with AaeUPO showing exceptionally higher total turnover number (200000) than MroUPO (26000) and rCciUPO (1400). Kinetic studies demonstrated that AaeUPO was the most efficient enzyme catalyzing stilbene dihydroxylation with catalytic efficiencies (k(cat)/K-m) one and two orders of magnitude higher than those of MroUPO and rCciUPO, so that 4-hydroxystilbene appears to be the best UPO substrate reported to date. In contrast, the peroxygenase from the ascomycete Chaetomium globosum (CglUPO) failed to hydroxylate trans-stilbene at the aromatic ring and instead produced the trans-epoxide in the alkenyl moiety. In addition, stilbenoids such as pinosylvin (Pin) and RSV were tested as substrates for the enzymatic synthesis of RSV from Pin and oxyresveratrol (oxyRSV) from both RSV and Pin. Overall, lower conversion rates and regioselectivities compared with trans-stilbene were accomplished by three of the UPOs, and no conversion was observed with CglUPO. The highest amount of RSV (63% of products) and oxyRSV (78%) were again attained with AaeUPO. True peroxygenase activity was demonstrated by incorporation of O-18 from (H2O2)-O-18 into the stilbene hydroxylation products. Differences in the number of phenylalanine residues at the heme access channels seems related to differences in aromatic hydroxylation activity, since they would facilitate substrate positioning by aromatic-aromatic interactions. The only ascomycete UPO tested (that of C. globosum) turned out to have the most differing active site (distal side of heme cavity) and reactivity with stilbenes resulting in ethenyl epoxidation instead of aromatic hydroxylation. The above oxyfunctionalizations by fungal UPOs represent a novel and simple alternative to chemical synthesis for the production of DHS, RSV and oxyRSV.

机译：这项工作给未指明的第一个证据peroxygenases(好的)担子菌里雨里地留在田头aegerita (AaeUPO) Coprinopsis灰质的特定选择的羟基化催化4-dihydroxy-trans-stilbene trans-stilbene 4(DHS),白藜芦醇(RSV)模拟的预防癌症入侵和影响转移最近被证明。完成转换的衬底(收益率DHS)是通过三个酶来完成的测试,使用过氧化氢作为唯一co-substrate AaeUPO总营业额显示异常高数量比MroUPO(200000)(26000)和rCciUPO(1400)。是最有效的酶催化对称二苯代乙烯dihydroxylation与催化效率(k (cat) / km)两个数量级高于MroUPO rCciUPO,所以4-hydroxystilbene似乎是最好的好的衬底报道日期。peroxygenase从子囊菌毛壳菌属globosum (CglUPO)未能羟化trans-stilbene芳环和代替生产中的trans-epoxide烯基一半。此外,种在如pinosylvin(销)和RSV的基板进行测试酶合成的RSV销oxyresveratrol oxyRSV RSV和销。总体而言,较低的转化率和区域选择性与trans-stilbene相比通过三个好的,不是吗转换与CglUPO观察。数量的RSV(63%的产品)和oxyRSV (78%)与AaeUPO再次获得。peroxygenase活动被证明了从(过氧化氢)-O-18 O-18整合到对称二苯代乙烯羟基化产品。血红素的苯丙氨酸残基的数量访问通道似乎相关的差异芳香族羟基化活动,因为他们会促进底物定位的aromatic-aromatic交互。子囊菌好的测试(c . globosum)原来最不同的活性部位(血红素腔远端)和反应对称二苯代乙烯乙烯环氧化作用而不是芳香族羟基化。oxyfunctionalizations真菌好的代表小说和简单的替代化学合成生产的国土安全部,RSV和

Selective synthesis of the resveratrol analogue 4,4 '-dihydroxy-trans-stilbene and stilbenoids modification by fungal peroxygenases

摘要

著录项

相似文献

相关主题

期刊订阅