| This paper studied on identification, mutagenesis and fermentation condition ofhigh-producing Hb proteinase strain. Investigated the pressing technology formultistage biodegradation of porcine blood. The research paper includes mainly fiveaspects:(1) Induced mutagenesis of high-producing Hb proteinase strain by DES andUltraviolet RayAs a original strain, the Bacillus pumilus strain Lact5.Ⅲ.6 was treated by DESand Ultraviolet (UV) twice. Mutagenesis and a mutant CN8 was obtained. The mutantCN8 of DH as 118.09%fold higher than the original strant was obtained under 1%(v/v) DES, for 30min, 30W UV lamp, at a distance 30cm from the protoplasts, for30s.Submerged culture showed that the hereditary character of high productivity ofmutant CN8 was stable.(2) Analysis of 16S rDNA sequence and FTIR spectroscopic of a high-yield Hbproteinase-pruducing strainA high-yield Hb proteinase-pruducing strain, which was isolated from soil, 16SrDNA was PCR and sequenced. The accession of GenBank is EF185299. The 16SrDNA phylogenetic trees was constructed by comparing with the published 16S rDNAsequences of the relative bacteria species. In the phylogenetic tree strain and Bacilluspumilus was the closest relative with 99.6-99.9% sequence similarity. According tothe phylogentic analysis it was identified as Bacillus pumilus.(3) Study on the mixed fermentation of porcine blood by multi-strainsSome particular cell components of intact Bacillus cereus could be detected andidentified by Fourier Transform infrared spectroscopy (FT-IR). Data were collecteddirectly, then through converting FTIR spectra of the samples into second derivativespectra. Typical marker bands were used to identify these bacterial cell componentssuch as capsules, endospores or storage materials. Capsules were detected in cell by astrong amide band near 1654cm-1 typical forα-helical structures and by strongcarboxylate stretching vibrations(≈1601 cm-1 and≈1403 cm-1, respectively).Formation of endospores was discovered using marker bands for dipicolinic acid(located at≈1617 cm-1,≈1372 cm-1 and≈1569 cm-1). Spectra of this strain showedexpression of poly-β-hydroxybutyric acid granules, capsules and endosporessimultaneously. These bacterial cell components can be identified by secondderivative FTIR spectroscopy which could distinguish the overlap spectrum. And thatoffered some referenced information for molecular biology and cellular biology.The purpose was to use porcine blood for the preparation of peptides in mixedmicroorganisms fermentation. The fermentative processe by mutation B.pumilus CN8, B.subtilis W11,N14, S. Cerevisiae were described. Various experimental conditions,such as inoculum size, time, temperature and cornmeal were investigated to optimizethe mixed fermentation. The result showed: the optimum mixed fermentationconditions were 2% CN8, 0.5% W11, 0.5% N14, 0.2% S.Cerevisiae inoculum size;33℃fermentation temperature, 60h fermentation time.(4) Study on the pressing technology for multistage biodegradation of porcinebloodThis research paper was studied on the influence that different process to themultistage biodegradation of porcine blood. The results were as following:hydrolyzing porcine blood through 40℃, pH=8.0, concentration of substrates 10%,enzyme quantity 8 000U/g, for 8h, then vacuum concentrated hydrolysis to substrates14%, then fermented the hydrolysis at 2% CN8, 0.5% W11, 0.5% N14 inoculum size,pH=7.25, at 33℃, for 60h.
|
PDF Full Text Request