A method for the bioaugmentation of vinegar by a protease-producing Streptomyces strain from marine sources
A bio-enhancing, streptomyces technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., to achieve the effect of improving the flavor and quality of vinegar and improving the utilization rate of raw materials
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Embodiment 1
[0031] Example 1: Screening of high-yield protease strains in marine sediments
[0032] (1) Pretreatment of marine sediment samples
[0033] The sediment was coated on the casein seawater plate by the direct dilution coating method, and cultured at 25°C for 20 days.
[0034] Seawater casein medium: NaH 2 PO 4 ·7H 2 O 1.07g, KH 2 PO 4 0.36g, casein 4g, agar 15g, aged sea water 1000mL.
[0035] (2) Initial screening of flat transparent circle
[0036] Select a single colony on the casein seawater plate and inoculate it on the screening medium, measure the diameter of the transparent circle (Rt) and the diameter of the colony (Rs) after culturing at 25°C for 48 hours, and calculate the circle diameter ratio K, K=Rt / Rs. Select a colony with a relatively large ring diameter for streaking and purification 2-3 times. After the microscopic examination confirms the pure culture, inoculate it on the slope and store it at 4°C, and store the bacterial suspension in 10% glycerol at ...
Embodiment 2
[0044] Example 2: Preparation of high protease-producing strain Streptomyces sp.CCTCC NO:M 2017210 lyophilized powder
[0045] (1) Activation culture of protease-producing strain Streptomyces sp.CCTCC NO:M 2017210
[0046] Put the bacterial powder of the strain Streptomyces sp.CCTCC NO:M 2017210 into 150mL liquid activation medium, and culture it on a shaker at 37°C for 48-50h to obtain the first-grade seed liquid; inoculate the first-grade seeds at 6% (v / v) Put the amount into 400mL liquid medium, and culture it on a shaker at 37°C for 48-50h to obtain the secondary seed liquid; put the secondary seed liquid into 6000mL liquid medium according to the inoculation amount of 8% (v / v), and shake it at 37°C Cultivate for 50-55h to obtain tertiary seed liquid.
[0047] Seawater LB medium: peptone 10g, yeast powder 5g, aged seawater 1000mL.
[0048] (2) Preparation of Streptomyces sp.CCTCC NO:M 2017210 lyophilized powder
[0049] Pour the tertiary fermentation broth into a 500mL ce...
Embodiment 3
[0050] Embodiment 3: the bioaugmentation of vinegar fermentation process
[0051] (1) Raw materials
[0052] The selected fermented vinegar raw material proportioning of the present embodiment is:
[0053] Wine mash: 130kg / cyl (alcohol content: 9.6, total acid: 4.4g / L); bran: 50kg / cyl; large bran: 24kg / cyl; water: 15kg / cyl; NaCl: 3kg / cyl.
[0054] (2) Traditional vinegar brewing process
[0055] [1] Add 130kg of wine mash and 50kg of bran into the cylinder, stir and mix.
[0056] [2] Add 8kg seed unstrained spirits in the cylinder.
[0057] [3] Heating stage: Rinse the fermented grains properly, add bran, and turn the fermented grains, but do not cut the bran down, and wait for the product temperature to rise above 40°C.
[0058] [4] Scooping stage: After the heating is completed, according to the product temperature, the bran is cut down, rinsed, added, and fermented every day. One or two days after the ladle starts, start to divide the vats. At this stage, the addition...
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