Malate dehydrogenase (EC 1.1,1,37.) (MDH) was purified to apparent homogeneity from the cytosolic fraction of the protozoan Trichomonas vaginalis Donné. The four step purification included ion-exchange chromatography (DEAE-Sephacel and Q-Sepharose, elution with NaCl) and affinity chromatography (Reactive Red Agarose, elution with NADH and NaCl). The enzyme was purified about 132-fold (30.6% yield) to a specific activity of 352 units mg~'. The Km values determined at pH 7.8 (pH optimum from 7.5 to 8.3) for oxaloacetate and NADH were 16.2 pM and 10.6 μΜ, respectively. The MDH activity was inhibited by the substrate, decreasing to 50% at about 1 mM concentration of oxaloacetate. The reverse reaction from malate to oxaloacetate showed a pH optimum around pH 9.5. The Km for malate and NAD* (determined at pH 7.8) were 1220 pM and 69.9 pM, respectively. SDS-PAGE analysis of the purified MDH revealed a single band with an apparent size of 34.5 kDa. The native molecular weight was estimated by HPLC gel filtration to be 60 kDa, which indicates that the T. vaginalis MDH exists as a dimer.
High level of phosphoenolpyruvate carboxylase (PEPC) gene was stably inherited and transferred from the male parent, PEPC transgenic rice, into a female parent, japonica rice cv. 9516. Relative to the female parent, the produced JAAS45 pollen lines exhibited high PEPC activity (17-fold increase) and also higher photosynthetic rates (about 36 %-fold increase). The JAAS45 pollen lines were more tolerant to photoinhibition and to photo-oxidative stress. Furthermore, JAAS45 pollen lines, as well as their male parent, were tested to exhibit a limiting C4 cycle by feeding with exogenous C4 primary products such as oxaloacetate (OAA). Thus the PEPC gene and photosynthetic characteristics of PEPC transgenic rice could be stably transferred to the hybrid progenies, which might open a new breeding approach to the integration of conventional hybridization and biological technology. and L. Ling, B. J. Zhang, D. M. Jiao.