Study On The Diversity And Metabolites Of Rhodotorula From The Offshore Areas In Leizhou Peninsula

Marine Rhodotorula from is a type of single-celled yeast strain which exist extensively in the oceans, which is rich in protein, unsaturated fatty, acid and essential amino-acid. It will produce a variety of active substances including carotenoid, digestive enzymes, vitamin, saccharides, oils and fats, and so on in the process of its metabolism.This article is to isolation and identification of Rhodotorula from the offshore areas in Leizhou peninsula, which provided some experimental data and basic theory of the development and application of animals Rhodotorula probiotics.We gathered samples including the marine organisms, seawater, sea mud and so on,from offshore areas in Leizhou peninsula, A total of 317 samples. First of all, sample enrichment culture, then isolated cultured and pure cultur by Improveing yeast agar plates and malt extract agar. Strains isolated by cell morphology characteristics, cultural characteristics, physiological and biochemical test results were preliminary identification;for preliminary identification of red yeast strains selected as representative 26 s r DNA D1/ D2 region of sequence analysis, phylogenetic tree, and diversity analysis. The results show that Isolation and identification of 71 marine Rhodotorula came from 317 samples.The 36 isolated strains of marine Rhodotorula yeast we selected as representative belonged to 5 species and 2 suspected new species. They were 22 isolated strains of the Rhodotorula mucilaginosa, 6 isolated strains of the Rhodotorula glutinis, 2isolated strains of the Rhodotorula slooffiae, 1 isolated strain of the Rhodotorula pallida,3 isolated strains of the Rhodosporidium and 2 isolated strain of a suspected new species,respectively named Rhodotorula. Sp. Nov. c ZDJ2 and Rhodotorula. Sp. Nov. ZTC2Cell walls of 71 marine Rhodotorula were treated separately by acid-heating dimethyl sulfoxide(DMSO) method, lactic acid extraction and freezing-thawing method.The purpose of obtaining bacterial cell extract, by spectrophotometric method quantitative determineing in bacterial cell extract of carotenoid content and qualitative analysising astaxanthin content, by HPLC quantitative detecting astaxanthin content of in bacterial cell extract of selected strains Acid-heating extraction was the best extraction method, DMSO screening technique Was the best screening technique. The content of the carotenoid of the 71 strains of the marine red yeast ranged from 54.127 ug/g to 608.428ug/g. The content of the carotenoid of the Rhodotorula mucilaginosa ZZC1 strain was the highest,it reached up to 608.428 μg/g. We screened out the strain which had the highest. The content of the astaxanthin of the Rhodotorula mucilaginosa ZTHY2 strain was the highest,it reached up to 11.641 μg/m L.We conducted a preliminary screening of the ability of protease, cellulose, amylase productivity of By hydrolysis of casein, sodium carboxymethylcellulose(CMC-Na), and starch to form a transparent circle method.We conducted secondary screening of the enzyme production strains by Folin development process, CMC saccharification force method, 3, 5-2 nitro salicylic acid method and on nitrophenol method. We screened out11 strains which could product protease, 16 strains which could product cellulose, 33 strainswhich could product amylase.71 strains which could product lipase. We found that the strain Rhodotorula mucilaginosa ZTHG5 had the highest protease productivity491.520 U, Rhodotorula mucilaginosa a ZDD6 had the highest cellulose productivity15.000 U, Rhodotorula mucilaginosa Rhodotorula mucilaginosa ZDHN1 had the highest amylase productivity 7.530 MMU, the strain Rhodotorula mucilaginosa ZTHY1 had the highest lipase productivity 6.209 U.By using of Vitamin B1, vitamin B2 and vitamin B6 standard calibration curve, by using of the ultraviolet spectrophotometry, we detected the content of vitamin B1 and vitamin B6 in 71 strains of red yeasts. Moreover, the content of vitamin B2 in 71 strains of red yeast was detected by visible spectrophotometry. The results showed that 71 strains Marine Rhodotorula produce vitamin B1, vitamin B2 and vitamin B6; we obtained that Rhodotorula mucilaginosa c ZDY2 had the highest content of vitamin B1 that is13.128 μg/m L, Rhodotorula mucilaginosa a ZDY2 strived the top place for vitamin B2 by14.711 μg/m L and Rhodotorula mucilaginosa c ZDD1 yielded highest vitamin B6 by9.660 μg/m L.Escherichia coli, Staphylococcus aureus, Salmonella pullorum and Bacillus subtilis ware selected as indicator bacteria. We used the filter method to study the bactweiostatic activity of strains of marine Rhodotorula. Eight strains had bacteriostatic activity in 71 strains of marine Rhodotorula. They are 2 strains of Rhodotorula mucilagiaosa, 3 strains of Rhodotorula slooffiae, 1 strains of Rhodotorula pallid, 2 strains suspected new species Rhodotorula. Sp. The maximum inhibition zone diameter only had 8.5 mm, less effective antibacterial activity.We separated 71 strains of marine Rhodotorula from the offshore areas in marine environment and marine organisms in Leizhou peninsula, Among them, Rhodotorula mucilaginosa and Rhodotorula glutinis are the dominant species. We selected outhigh-yielding carotenoids, astaxanthin, protease, cellulase, amylase, lipase, vitamin B1,vitamin B2 and vitamin B6 strain, But not easy to produce antibacterial substances.