Vestavia Hills, AL

来源: 于 2020-11-24 09:23:03 [博客] [旧帖] [给我悄悄话] 本文已被阅读：次

Run-Tao Yan

Research Associate at University of Alabama at Birmingham · Birmingham
   (205) 325-8617
rtyan@uab.edu
yan@uab.edu

1201 11th Ave S # 400

 Birmingham

 Alabama

 35205-3410

 http://www.uab.edu

 

  
Mobile number

(917) 923-4438

 

Born

March 26, 1956

Marital status

Single

2800 Crossgate Cir, Vestavia Hills, AL, 35216-3177

 

 

 

 

 

 
 
 
 
 
 

Enzmology of butanol formation in Clostridium Beijerinckii

Yan, Run-Tao (Virginia Tech, 1991).

The present study encompasses an investigation of the expression of solvent forming enzymes and purification and characterization of butanol-forming enzymes. More sensitive and accurate procedures for the determination of acids and solvents in cultures have been developed, which led to the recognition of the onset of solvent production at the mid-exponential phase, about two h earlier than previously reported. Activities of solvent-forming enzymes started to increase about one h before the onset of measurable solvent production and the activities of solvent-forming enzymes did not increase simultaneously. CoA-acylating aldehyde dehydrogenase (ALDH) was purified to near homogeneity. The ALDH showed a native M.. of 100,000, and a subunit Mr of 55,000. ALDH could use either NAD(H) or NADP(H) as the coenzyme. ALDH was oxygenlabile. The O2-inactivated enzyme could be reactivated by incubating the enzyme with CoA. Both NADH- and NADPH-dependent alcohol dehydrogenase activities were present in crude extracts. The ratio of NADPH-dependent activity to NADH-dependent activity (the PID ratio) varied in crude extracts. The PID ratio was affected by O~ ionic strength, pH, growth stage of cell, Fe in culture medium and temperature. Two ADHs have been identified in crude extracts. The NADPH-dependent ADH (P-ADH) could be separated from the NADH/NADPH-dependent ADH (D/P-ADH). The D/P-ADH has been extensively purified. The D/P-ADH showed a native Mr of 70,000 and subunits with Mr of 45,300 and 40,000. The D/P-ADH activity could be inactivated by a,a' -dipyridyl and restored by Fe2+.