Bacillus amyloliquefaciens and application thereof in solid-state fermentation vinegar brewing
A technology for dissolving starch spores and solid-state fermentation, which is applied in the preparation of vinegar, bacteria, food science, etc., to achieve the effects of increasing the rate of vinegar production, increasing the content of ammonia nitrogen, and reducing production costs
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[0035] Example 1:
[0036] Screening of ferulate esterase producing bacteria and determination of enzyme activity:
[0037] From the solid-state vinegar fermentation tank in the natural fermentation state of Qianhe Weiye Food Co., Ltd., the vinegar fermented grains on the 2nd, 4th, 6th, 8th, 10th, 12th, 14th, 6th, 18th, 20th, 22nd and 24th day of the fermentation cycle are selected respectively. The sampling method is to take vertical samples from the surface to the bottom of the vinegar fermented grains around the fermentation tank, and then evenly mix the vinegar fermented grains samples in different fermentation cycles to obtain strain screening samples. Specific methods of screening:
[0038] Weigh 100g vinegar fermented grains and put them into 1000ml 0.85% sterilized normal saline, stir evenly and then stand still. Take 10ml supernatant into 100ml enrichment medium and culture overnight at 30 ℃ and 150rpm to obtain the primary culture solution. Continue to take 10ml of the o...
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[0046] Example 2:
[0047] Re screening of ferulic acid decarboxylase producing bacteria:
[0048] The ferulate esterase producing bacteria screened in example 1 were inoculated into the screening medium, cultured at 37 ℃ and 150 rpm for 3 days, and the supernatant was taken to detect the contents of 4-vinyl guaiacol and 4-ethyl guaiacol. The higher the content of 4-vinyl guaiacol and 4-ethyl guaiacol, the higher the activity of ferulic acid decarboxylase. The screening medium is: ferulic acid 2G / L, glucose 10g / L, yeast powder 2G / L, (NH 4 ) 2 SO 4 1.2g / L,KH 2 PO 4 0.37g / L,MgSO 4 ·7H 2 O 0.25g / L,CaCl 2 ·2H 2 O 0.05g / l, distilled water 1000ml, pH natural. 4-vinyl guaiacol and 4-ethyl guaiacol were determined by GC-MS. the fermentation supernatant was collected by centrifugation, 5 times the volume of dichloromethane and a certain amount of 2-octanol (internal standard) were added, and the organic phase was obtained after ultrasonic extraction for 15min, and the extraction was r...
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[0053] Example 3:
[0054] Identification of strain qh-20002:
[0055] 1. Morphological identification:
[0056] The strain qh-20002 screened in example 1 was inoculated on the solid medium and cultured at 37 ℃ for 24 hours to form an irregular shape, raised in the middle, containing mucus, serrated milky white colony with a diameter of 1-4mm. Gram staining observation: pink short rod, the colony morphology of amylolytic Bacillus qh-20002 is as follows: Figure 1 As shown in, the evolutionary tree analysis of strain qh-20002 based on 16S rDNA is as follows: Figure 2 As shown in.
[0057] Solid medium composition: sodium chloride 10g / L, peptone 10g / L, yeast powder 5g / L, agar 20g / L, and the solvent is deionized water.
[0058] 2. Physiological and biochemical identification:
[0059] 94 phenotypes of strain qh-20002 were tested by Biolog (Gen Ⅲ) automatic microbial identification system, including 71 carbon source utilization tests and 23 chemical sensitivity tests: strain qh...
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