| The use of marine microbial resources and exploitation has become a hot area of research at home and abroad. By 2000, worldwide, from the marine flora and fauna and micro-organisms isolated a new compound of 10,000 species, of which patent applications have more than 200 kinds of chemical compounds.These studies greatly facilitate and promote the study of marine micro-organisms, so that marine micro-organisms into the marine pharmaceutical and biotechnology in international hot spots and front.Carotenoids are a class of C40 terpenoid compounds and their derivatives in general, from eight isoprenoid units, yellow, orange-red or red pigment. Biological functions of carotenoids rich in vitamin A as it is the original, can prevent night blindness, can be anti-oxidant, anti-cancer, coloring and strong increase of the gap junction intercellular communication,, can enhance the animal’s reproductive capability.The first chapter reviews the status of the development of marine microbial resources; carotenoid structure; the nature of the carotenoid; carotenoid physiological activity; natural carotenoid biosynthesis; the analysis of carotenoid identification; bacteria and fermentation physiological conditions; the purpose of the study and significance.Chapter II study from the East China Sea (north latitude, 31.237 o, longitude 124.605 o) 160 meters deep to obtain sediment samples collected, diluted coating, observed colony color, initially with five strains produced a red pigment, selecting one of the most red, a color strain, after extraction of pigment extraction, the use of HPLC and UV detection of full spectrum scanning, to determine its formation nearly 10 kinds of natural carotenoids. Subsequently the use of morphological, physiological, biochemical, whole-cell fatty acid detection and taxonomy of 16S rDNA and many other technologies, taxonomic identification of the strains and to identify it as Kocuria genus, named as Koura sp.Car-red.Chapter III discusses the use of microwave method, ultrasonic method and the HCl method of the broken strain Car-red effect. According to the experimental results, HCl’s broken the most ideal results, the best extraction conditions HCl5mol / l concentration of extraction under the conditions of 120min, extraction of 0.409μg/mL. And discussed in the broken after the choice of extraction solvent, after repeated experiments, found that ethanol extract the best, can be achieved 0.526μg/mL. Finally the orthogonal experimental design methods to broken time, extraction temperature, extraction time, extraction solution volume was 4 factors with 3 levels of extraction condition optimization experiments, the optimal extraction conditions has been a priority level in accordance with its combination of carotenoid yield an average of up to 0.693μg/mL. The fourth chapter from the culture medium and fermentation conditions optimization formula optimization to improve both the strains of Car-red total carotenoid production. Single factor and orthogonal design experiments to find the final optimized medium formula, that is, the amount of sucrose by adding 0.5%, by adding the amount of 0.75% peptone, yeast extract by adding the amount of 0.25%, magnesium sulfate by adding the amount of 0.15%. Theoretical yield can be achieved 1.160μg/mL, through the verification experiment, the actual production of 1.139μg/mL. The liquid volume 20mL, inoculum 12%, the growth temperature of 28℃, shaking speed 120 r / min for the more ideal fermentation conditions.Chapter V main research contents for the use of PCR technology to strain Car-red as a template, amplification of the strain CrtB genes, and outgoing sequence, according to sequencing results to obtain a size of 578 bp sequence of the gene sequencing results will be logged NCBI’s GenBank on the sequence homology comparison of the results showed that: The existence of strains with the carotenoid biosynthesis-related genes CrtB. From the genetic point of view is verified strain Car-red is indeed produced carotenoids of marine strains. |
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