Influence Of Different Cultivate Conditions On Yield Of Conjugated Linoleic Acid And The MRNA Expressed Level Of LAI Gene In Lactobacillus Plantarum P8

Conjugated Linoleic Acid (CLA) is a natural fatty acid which have kinds of bioactive and exists differential in geometry configuration and conjugated double bond at octadecadienoic acid.Recently researches found that, as a unstaturated fatty acid, CLA possessed many physiologic function such as canticancer, proved human’s immunity, increased muscles, cut down the level of the fat and the cholest, rosed the density of bones, rejects the artheroschlerosis. the CLA palied a very important role in medical domain, foods, cosmetics and bait vessel industry. An important sense is that we make many researches on LAI as a key enzyme in the transfer of the LA to CLA.So far, in the aspects of the structure, function, property and regulation of LAI, people often optimized the LAI by the technique of gene engineering and biotechnology. At present, many papers indicated that microbes could promoted the process of the LA to CLA as a narrow spectrum by the LAI. The biosynthesis is very few, and this would become the future develop tendency in this area.This paper put the Lactobacillus plantarum P8as our materials. The measurement target was the cultivante time, the contens of inoculation, carbon sources, nitrogen sources, the concentration of the LA, phvolue, cultivante temperature at el. Through the single factor analyze and the orthogonal experiment, the optimization conditions were definited for how to cultivate the Lactobacillus plantarum P8, that the cultivante time was22h, the contens of inoculation is1.5%, carbon sources is the amylaceum which the concertration was3g/L, nitrogen sources was the tryptone which the concertration was2g/L, the concentration of the LA was0.9mg/μL. Under this condition, the maximum daily yield of CLA in Lactobacillus plantarum P8could reach0.0692mg/mL.N-hexane extraction, absorption spectrometry, methods of Trizol, reverse transcription PCR, real-time PCR were used to research the mRNA expressed level of LAI gene in Lactobacillus plantarum P8. Total RNA of16different cutivated groups were abstracted. And agarose gel electrophoresis and absorption spectrometry were us(?) to detection the integration and purity of RNA, the ratio of OD260/OD280between1.90-2.10, thus were completely suit RT-PCR, real-time PCR.Specific primer were designed and were used to reverse transcription PCR, and in progress of Real-time PCR to detected the mRNA expressed level of16different cultivated groups. This paper indicated that, LAI gene expressed in all the groups, but the expressed level were different. The lower level groups is the4th,6th and12th groups. And in the groups of the1th,5th,10th, the mRNA expressed level is much higher. These indicated that different cultivation is influenced the mRNA expressed levels in P8.