Cordysobin, a novel alkaline serine protease with HIV-1 reverse transcriptase inhibitory activity from the medicinal mushroom Cordyceps sobolifera.
A novel serine protease, designated as cordysobin, was purified from dried fruiting bodies of the mushroom Cordyceps sobolifera. The isolation procedure utilized ion exchange chromatography on DEAE-cellulose and SP-Sepharose followed by gel filtration on Superdex 75. The protease did not adsorb on DEAE-cellulose but bound to SP-Sepharose. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the protease resolved as a single band with an apparent molecular mass of 31 kDa. Its optimal pH was 10.0, and the optimal temperature was 65°C. The protease displayed a K(m) value of 0.41 μM and 13.44 μM·min⁻¹ using Suc-Leu-Leu-Val-Tyr-MCA as substrate at pH 10.0 and 37°C. Protease activity was enhanced by the Fe²⁺ ion at low concentration range of 1.25-10 mM and was strongly inhibited by Hg²⁺ up to 1.25 mM. The protease was strongly inhibited by chymostatin and phenylmethylsulfonyl fluoride (PMSF), suggesting that it is a serine protease. It manifested significant inhibitory activity toward HIV-1 reverse transcriptase (RT) with an IC₅₀ value of 8.2×10⁻³ μM, which is the highest anti-HIV-1 RT activity of reported mushroom proteins.
State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing 100193, China.
Shou-Xian
SX
Wang
Yu
Y
Liu
Guo-Qing
GQ
Zhang
Shuang
S
Zhao
Feng
F
Xu
Xiao-Li
XL
Geng
He-Xiang
HX
Wang
D019380:Anti-HIV Agents / Q000302:isolation & purification / Q000494:pharmacology*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D032661:Cordyceps / Q000201:enzymology*; D003374:Coumarins; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D048690:Fruiting Bodies, Fungal / Q000201:enzymology; D005656:Fungal Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D054303:HIV Reverse Transcriptase / Q000037:antagonists & inhibitors*; D015497:HIV-1 / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D020128:Inhibitory Concentration 50; D013058:Mass Spectrometry; D008970:Molecular Weight; D009842:Oligopeptides; D057057:Serine Proteases / Q000302:isolation & purification / Q000494:pharmacology*; D013696:Temperature
D019380:Anti-HIV Agents; D003374:Coumarins; D005656:Fungal Proteins; D009842:Oligopeptides; C046582:succinyl-leucyl-leucyl-valyl-tyrosyl-methylcoumarinamide; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase; D057057:Serine Proteases; C571830:cordysobin protein, Cordyceps sobolifera