TH112259A - "Rapid enzyme detection and screening process" - Google Patents

Abstract

------ 31/07/2018 ------ (OCR) Page 1 of 1 page Summary of Invention This invention focuses on the development of enzyme detection and screening processes. The methods for screening enzymes from enzyme-producing bacteria were improved and stored in cells. Normally, enzymes that are produced and accumulated in cells take quite a long time before enzymes are released outside the cells by leaking the enzymes through the bacterial cell wall. There is a leak and the enzyme is released without affecting the cell growth by a method known as the pouring technique. Which will mix various substances That affects bacterial cell wall leakage, causing bacteria to release more and more enzymes, enabling enzyme activity to be monitored with higher sensitivities and less time. The invention used a cheaper chemical to induce a leak in the cell wall, instead of using the same substance with agar-over-agar-over-cultured bacteria technique. The ability of chemicals to cause leakage of the cell wall was tested. For application in the measurement of the activity of the enzyme 'polysaccharide', polysaccharide, lipase and bacterial protease ------------ This invention has focused on Focus on the development of processes for screening and screening various enzymes. Quickly by improving Method for screening for enzymes from bacteria that produce enzymes and stored in the cell. Which is usually the enzyme that is produced and accumulated In cells, it takes quite a long time for the enzyme to be released outside the cell by leaking the enzyme through the bacterial cell wall.Therefore, methods have been tried to leak the bacterial cell wall and release the enzyme by: Without Effect on cell growth by a method known as the pouring technique. Which will mix various substances Affecting the leakage of the wall Becteria cells allow the becteria to release more and more enzymes that can be examined. Enzyme activity can be achieved with higher sensing sensitivity levels and less time consuming. This invention uses chemicals that are It is cheaper to induce a leak in the cell wall instead of using the same substance in conjunction with the agar-over pouring technique. Cultured on solid food The ability of chemicals to induce leakage of the cell wall is performed for application in the determination of enzyme activity in the bacterial polysaccharide, lipase and protinase subgroups.

Claims (8)

Disclaimer (all) which will not appear on the advertisement page. : ------ 31/07/2018 ------ (OCR) Page 1 of 2 pages Disclaimer 1. Rapid enzyme detection and screening process. With the following steps: 1.1 Preparation of solid culture medium to be mixed with enzyme substrate to be tested and screened 1.2 Cultivation of enzyme-producing bacterial cells to be examined and screened on solid food 1.3 The colonization of enzyme-producing bacteria to be examined and screened with chemical agar agar can leak bacterial cells. The chemicals that Mirit Kang can choose from EDTA, ampicisin, sodium dodexyl sulfate, zacosine or triton xl -! 00 One or more combinations 1.4 Examination of the enzyme activity is characterized by The enzyme-induced leaks in the cell wall to release the enzyme together with the colonization technique of enzyme-producing bacteria to screen the enzyme on solid food. It can monitor the activity of the enzyme sweeping and moistening. 2. Rapid enzyme detection and screening process. According to claim 1, where the solid culture media mixed with such a substrate Its characteristics consist of 0.05% of substrates for screening enzyme-producing bacteria, 1% tributyrate, 0.1% gum aerobic or 2% skim milk. Rapid screening of enzymes According to claim 1, where the culture of enzyme-producing bacterial cells to be tested and screened on solid food There is a special feature The cells were cultured at the appropriate temperature for 16 - 48 h, then the cells were reduced to 5 - 20 coloni cells per plate. Then spread on solid food with the said chop And cultivate such cells at the right temperature Until they grow into colony. 4. Process for enzyme detection and screening quickly. According to the 1st claim, where the colonization of enzyme-producing bacteria The special feature is Pour the bacteria's colony with 0.7% agar mixed with a chemical that "makes" the bacteria cells to leak. Where selectable from EDTA at a concentration of 10 to 50 mmolary, an ampicillin at a concentration of 25 to 50 mmol, a sodium dodecyl sulfate (SDS) at a concentration of 0.1% to 1.0%, zacosine at a concentration of 0.1% to 0.5%, or Triton XL -! 00 The concentration of 0.05 to 0.1 percent. 5. Process of enzyme detection and screening rapidly. According to either claim 1 or 4, where the pouring of \ 'co \' loni of the enzyme-producing bacteria Has the characteristics of The bacteria's colonies are poured with 0.7% agar mixed with chemicals that have the potential to leak more than one bacterial cell. Selectable from (5.1) Ampicisin and EDTA (5.2) RAMPicillin and SDS (5.3) Ampisin and Tritonxl -! 00 (5.4) Ampicisin and zacosine (5.5), SDS and zacosine (5.6), SDS and tritonexl-P 0 (5.7), zacosine and triton xl. -! 00 or (5.8) SDS, zacosine and triton xl -! 00 6. Rapid enzyme screening and screening process. Per claim 1, 4 or 5, where the colonization of enzyme-producing bacteria There is a special feature The colony of the bacteria is poured with 0.7% agar mixed with insoluble strate onto the colonic growing on the plate. Then cultured the culture that needs to be examined at 37 degrees Celsius for 1 hour. 7. Process of enzyme detection and screening quickly. According to one of the claims 1, 4, 5 or 6, where the colonization of enzyme-producing bacteria The special feature is Pour the colonic of the bacteria with 0.7% agar in a buffer solution at pH 3 to 10 in order to screen for the enzyme that works well in the strongly acidic environment. Page 2 of the number 2 page 8. Method of detection. And rapidly screening enzymes According to one of the claims 1, 4, 5, 6 or 7, where the colonization of enzyme-producing bacteria There is a special feature The bacterial colonies are poured with 0.7% agar mixed with more than one strate to detect the activity of more than one enzyme at the same time ------------ 1. Rapid enzyme detection and screening process The steps are as follows: 1.1 Preparation of solid culture media that is mixed with the enzyme sludge to be examined and screened. 1.2 Cultivation of enzyme-producing bacteria cells to be examined and screened on solid food 1.3. Of bacteria producing enzymes that need to be examined and screened with agar that is mixed with chemicals that have Bacterial cells can leak and 1.4 enzyme activity monitoring. With special characteristics Causing a leak in the cell wall to release enzymes Together with the technique of pouring colonies Of bacteria that produce enzymes To screen enzymes on solid foods Which can monitor the work activity of Enzymes can be clearer and faster. 2. Rapid enzyme detection and screening process According to claim 1, where the solid culture media is mixed Substrate it It is characterized by having 0.05% of the substrate for screening enzyme-producing bacteria. Tributyrin 1%, Gum Aerobic 0.1% or Skim Milk 2% 3. Rapid enzyme detection and screening process According to claim 1, where the bacterial cell culture produced Enzymes to be examined and screened on solid food There is a special feature The cells were cultured at the appropriate temperature for approximately 16-48 hours, then the cells were reduced to 5-20 colonies per plate and spread on solid food containing the substrate. And cultivate such cells at the right temperature Until growing up into a colony 4. Rapid enzyme detection and screening process According to claim 1, where the colonization of enzyme-producing bacteria has special characteristics: The colonies of the bacteria are poured with 0.7% agar mixed with chemicals that can cause cells. Leaking bacteria Where selectable from EDTA at a concentration of approximately 10 to 50 mmolar; ampicillin at the Concentration of approximately 25 to 50 mmol, sodium dodexyl sulfate (SDS) at a concentration of approximately 0.1% to 1.0, zacosine at a concentration of approximately 0.1 to 0.5%, or Triton X-100. At a concentration of approximately 0.05% to 0.1 5. Rapid enzyme detection and screening process According to claims 1 or 4, any of the above. Of enzyme-producing bacteria There is a special feature The bacterial colonies are poured with 0.7% agar mixed with a chemical Has the effect of causing more than one type of bacterial cells to leak Selectable from (5.1) Ampicillin and EDTA (5.2) Ampicillin and SDS (5.3) Ampicillin and Tritonx-100 ( 5.7) Sacosine and Triton X-100 or (5.8) SDS, Sacosine and Triton X-100 6. Rapid enzyme examination and screening process Per claim 1, 4 or 5, where the colonization of enzyme-producing bacteria There is a special feature The bacteria colonies are poured with 0.7% agar mixed with insoluble substrate onto the colonies growing on the plates. And then culturing the bacteria to be examined at 37 degrees Celsius for about 1 hour 7. Rapid enzyme detection and screening process According to one of the claims 1, 4, 5 or 6, where the colonization of enzyme-producing bacteria There is a special feature Bacterial colonies are poured with 0.7% agar in a buffer solution at pH3 to 10 for screening of enzymes that perform well in strong acidic conditions. 8. Rapid enzyme detection and screening process According to one of the claims 1, 4, 5, 6 or 7, where the colonization of enzyme-producing bacteria There is a special feature The bacterial colonies were tapped with 0.7% agar mixed with more than one substrate to detect the activity of more than one enzyme at the same time.