The process of producing a protease enzyme specific for the degradation of silk glue protein from bacteria.
Patent Information
- Application Number
- TH2301005524
- Authority / Receiving Office
- TH · TH
- Patent Type
- Applications
- Current Assignee / Owner
- Filing Date
- 2023-09-05
- Publication Date
- 2026-08-31
Smart Images
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Abstract
Claims
OCR10KL (10 / 08 / 2569) 1. The process of producing protease enzymes specific for the degradation of silk glue protein from bacteria involves the following steps: a. Culturing bacterial seed cultures with a specific nutrient formula for cell growth, incubated at 25-35 degrees Celsius for 24-48 hours under shaking conditions at a speed of 150-250 rpm. b. Culturing bacteria to stimulate the cells to produce protease enzymes specific to silk glue protein but without damaging the silk fibers in a fermentation tank with a bacterial culture formula containing a specific ratio... The carbon-to-nitrogen ratio to inducing agent was in the range of 3:5:0.3 to 17:25:0.7 by mass / volume. Specific protease enzymes for silk glue protein but without damaging silk fibers were harvested from fermentation broth and extracted from cell sludge using a surfactant solution. The obtained enzymes were then mixed. The process of producing protease enzymes specific for silk glue protein degradation from bacteria according to claim 1, where the selected bacteria are Bacillus spp., Arthrobacters pp., Streptomycess pp., and Flavobacterium spp.The process of producing a protease enzyme specific for the hydrolysis of silk glue protein from bacteria according to claim1, where the most suitable bacteria is Bacillus sp. 16-14. The process of producing a protease enzyme specific for the hydrolysis of silk glue protein from bacteria according to claim1, where the specific bacterial culture medium for cell growth is a liquid BMSMS medium consisting of yeast extract 5-20 g / L of liquid BMSMS, glucose 10-40 g / L of liquid BMSMS, dipotassium hydrogen phosphate 0.5-2.0 g / L of liquid BMSMS, magnesium sulfate heptahydrate 0.1-0.3 g / L of liquid BMSMS, and silk glue protein 2-10 g / L of liquid BMSMS.The process of producing a protease enzyme specific for the hydrolysis of silk glue protein from bacteria according to claim 1, where the bacterial cultures are induced to produce a protease enzyme specific to silk glue protein but do not damage the silk fibers in the aforementioned fermentation tank, with an optimal bacterial count in the range of 5 to 10 percent by volume.
6. The process of producing a protease enzyme specific for the hydrolysis of silk glue protein from bacteria according to claim 5, where the optimal bacterial count has a cell concentration measured by the absorbance at a wavelength of 600 nm in the range of 0.1-2.0 per milliliter.
7. The process of producing a protease enzyme specific for the hydrolysis of silk glue protein from bacteria according to claim 1, where the carbon source can be selected from mashed soybean meal, molasses, cassava starch, and mung bean meal, either individually or in combination.
8. The process of producing a protease enzyme specific for the hydrolysis of silk glue protein from bacteria according to claim 1, where the nitrogen source can be selected from yeast extract, whey, and soy protein, either individually or in combination. 9.
10. The process of producing a protease enzyme specific for the hydrolysis of silk glue protein from bacteria according to claim 1, where the inducing agent is selectable from either sericin protein or a combination of both, and raw silk powder.
11. The process of producing a protease enzyme specific for the hydrolysis of silk glue protein from bacteria according to claim 1, where the optimal temperature for culturing bacteria to stimulate cell production of protease enzyme specific to silk glue protein without damaging the silk fibers in the fermentation tank is 25-35 degrees Celsius.
12. The process of producing a protease enzyme specific for the hydrolysis of silk glue protein from bacteria according to claim 1, where the optimal stirring rate for culturing bacteria to stimulate cell production of protease enzyme specific to silk glue protein without damaging the silk fibers in the fermentation tank is in the range of 400 to 600 revolutions per minute.
13. The process of producing a protease enzyme specific for the hydrolysis of silk glue protein from bacteria according to claim 1, where the optimal time for culturing bacteria to stimulate the cells to produce a protease enzyme specific to silk glue protein without damaging the silk fibers in a fermentation tank is between 24 and 72 hours.
14. The process of producing a protease enzyme specific for the hydrolysis of silk glue protein from bacteria according to claim 1, where the extraction of the residual protease enzyme in the sludge using the aforementioned surfactant solution has a ratio between the weight of the sludge and the volume of the surfactant solution in the range of 1:1 to 1:3.
15. The process of producing a protease enzyme specific for the hydrolysis of silk glue protein from bacteria according to claim 1, where the extraction of the residual protease enzyme in the sludge using the aforementioned surfactant solution has an optimal temperature range of 4-16 degrees Celsius.16.
15. The process of producing a protease enzyme specific for the hydrolysis of bacterial silk glue protein according to claim 1, where the extraction of residual protease enzyme in sludge using the aforementioned surfactant solution has an optimal stirring speed in the range of 500-2,000 rpm.
16. The process of producing a protease enzyme specific for the hydrolysis of bacterial silk glue protein according to claim 1, where the extraction of residual protease enzyme in sludge using the aforementioned surfactant solution has an optimal stirring time in the range of 30-180 minutes.
17. The process of producing a protease enzyme specific for the hydrolysis of bacterial silk glue protein according to claim 1, where the surfactant can be selected from 4-octylphenol polyate thoxylate and polysorbate 80, either individually or in combination.
18. The process of producing a protease enzyme specific for the hydrolysis of bacterial silk glue protein according to claim 1 or 17, where the most suitable surfactant is polysorbate 80. 19.The process of producing a protease enzyme specific for the degradation of bacterial silk glue protein according to patent18, where the polysorbate solution 80 has an optimal concentration in the range of 0.1% to 0.5% by mass per volume;