Production method of erythritol, variant strain of P. spp. and use thereof
A technology of erythritol and strains, applied in the field of biocatalyst compositions for erythritol manufacturing, can solve the problems of safety, poor industrial economy, and unrealized industrialization.
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Embodiment 1
[0081] Example 1. Isolation of Saccharomyces spp. SYC-ERY1 mother strain
[0082] The microorganisms used in the present invention were isolated according to the following method. The honeycomb was cultured in a liquid medium containing 35 (w / v)% glucose and 1.5 (w / v)% yeast extract under the conditions of 30 degrees and 250 rpm for 6 days. The cultivated culture solution was spread on a solid medium of the same composition to obtain about 5000 colonies.
[0083] The colonies obtained by strain isolation were inoculated in a medium containing 45(w / v)% glucose, 0.5(w / v)% yeast extract, 0.35(w / v)% KNO 3 , 0.025(w / v)%KH 2 PO 4 , 5ppm MnSO 4 , 3 ppm Thiamine (Thiamine) in 4 ml (test tube) culture medium (pH 5.5), and cultured at 30° C. and 200 rpm for 6 days.
[0084] The culture solution cultured for 6 days was centrifuged at 13,000 rpm for 10 minutes to remove the cells to obtain only the culture supernatant, and the supernatant was analyzed by thin layer chromatography (...
Embodiment 2
[0091] The characteristics of the bacterial strain screened in embodiment 2. embodiment 1
[0092] In order to understand the characteristics of the bacterial strains screened in the above-mentioned Example 1 and to identify bacterial strains, to the Korean Collection for Type cultures (KCTC) of Korea Research institute of Bioscience and Biotechnology (Korean Collection for Type Cultures of Korea Research Institute of Bioscience and Biotechnology Institute, located in South Korea No. 125 Keke Road, Rucheng District, Daejeon) to apply for and conduct identification.
[0093] The physiological and biochemical characteristics of this strain are as follows:
[0094] 1) Assimilation of sugar and organic acid
[0095] Soluble starch(+)
[0097] L-arabinose (-)
[0098] Glycerol (+)
[0099] 2) Base sequence analysis of D1 / D2 and ITS region of 26S rRNA gene
[0100] It was found that they were the same strain because the sequence similarity with...
experiment example 1
[0104] * Experimental example 1. Screening mutant strains and confirming erythritol production
[0105] Mutant strains were isolated by subjecting the strains (mother strains) screened in Example 1 to ultraviolet mutation (UV mutation). Specifically, the bacterial strain (wild type) screened in Example 1 was cultured for 2 days in 2 (w / v)% glucose, 2 (w / v)% malt extract, 0.1 (w / v)% peptone medium , using the culture medium to carry out ultraviolet mutation. In the ultraviolet mutation, the bacterial culture solution was diluted and applied to the ultraviolet mutation medium (45 (w / v)% glucose, yeast nitrogen source). After the smeared plate was irradiated with ultraviolet rays for 5 minutes under carcinogenic conditions, it was wrapped with foil and incubated in a 30° C. incubator to obtain colonies. The obtained colony was cultured in the same manner as when the strain was isolated, and subjected to TLC analysis and HPLC RI analysis, and cultured in a 250 ml flask (50 ml ...
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