FIG. 3.

Conversion of VHOH to VB and conversion of OAVN to AVR by the recombinant VHOH cyclase expressed in P. pastoris. VHA (A) or OAVN (B) was incubated with cell extract of P. pastoris. In panel A, porcine esterase was also added in order to convert VHA to VHOH for determination of the VHOH enzyme activity. The reaction products were extracted with chloroform and analyzed by TLC. Lane 1, no cell extract; lane 2, cell extract of P. pastoris transformed with only the vector; lane 3, cell extract of P. pastoris transformed with the vector containing the vbs gene. The small amount of VHOH in panel A was due to the low extraction efficiency of the high-polarity substance VHOH with chloroform.