Manipulating pyruvate decarboxylase by addition of enzyme regulators during fermentation of Rhizopus oryzae to enhance lactic acid production.

Ethanol was found as the major by-product in lactate fermentation by Rhizopus oryzae. Several methods have been conducted in order to limit ethanol formation, thus increasing the lactate yield. The direct way to suppress ethanol production can be done by inhibition of the responsible enzymes in the related pathway. Pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) are responsible for ethanol production in R. oryzae. Shunting the ethanol production pathway by targeting at PDC was attempted in this study. Three compounds including 4-methylpyrazole, glyoxylic acid, and 3-hydroxypyruvate with the in vitro reversible inhibitory effect on PDC were selected from the literature and were used to regulate the living cell of R. oryzae during the fermentation. The results show that 0.1 mM 4-methylpyrazole of which the structure resembled a thiazolium ring in thiamine diphosphate, PDC cofactor, and 1.0 µm 3-hydroxypyruvate, pyruvate analog, effectively hampered ethanol production. Further observation on the enzyme expression indicated that these two regulators not only targeted PDC but also caused changes in ADH and lactate dehydrogenase (LDH) activities. This was perhaps due to the living cell of R. oryzae that responded to the presence of the regulators to balance the pyruvate flux and subsequently maintain its metabolic activities.

Rapidly stopping hemorrhage by enhancing blood clotting at an opened wound using chitosan/polylactic acid/polycaprolactone wound dressing device.

Doxycycline and monosodium glutamate (MSG) loaded chitosan (CHI)/polylactic acid (PLA)/polycaprolactone (PCL) blend film was studied as a model device to deliver drug to targeted human organ which in this case was the skin with opened wound. The CHI/PLA/PCL blend film containing 60 % CHI, 28 % PLA, and 12 % PCL exhibited the good properties for making the dressing device. It was observed that doxycycline/MSG loaded CHI/PLA/PCL blend film could rapidly deliver both doxycycline and MSG at the high release percentage approaching 100 % loaded. MSG accelerated blood clotting and fibrin formation; thus, it exhibited the good hemostatic activity. The antibacterial activity of doxycycline loaded CHI/PLA/PCL blend film against Staphylococcus aureus and Escherichia coli as model bacteria was investigated. Doxycycline release played the crucial role in bacterial inhibition as observed from the lowest bacterial cell dry weight observed when compared with the control bacterial culture or the bacterial cultures with the presence of other films studied.