CN104263662A - Broad-spectrum antibacterial microzyme and method for enhancing antibacterial effect thereof - Google Patents
Broad-spectrum antibacterial microzyme and method for enhancing antibacterial effect thereof Download PDFInfo
- Publication number
- CN104263662A CN104263662A CN201410449239.7A CN201410449239A CN104263662A CN 104263662 A CN104263662 A CN 104263662A CN 201410449239 A CN201410449239 A CN 201410449239A CN 104263662 A CN104263662 A CN 104263662A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- furnished
- dare
- torulaspora delbrueckii
- subordinate phase
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
Abstract
The invention relates to a microzyme and a method for enhancing antibacterial effect thereof, particularly a broad-spectrum antibacterial microzyme and a method for enhancing antibacterial effect thereof. The microzyme is Torulaspora delbrueckii M8 which is collected by China Center for Type Culture Collection, and the collection number is CCTCC NO:M2013277. The method for enhancing antibacterial effect is implemented by carrying out fermentation culture on the Torulaspora delbrueckii M8 with a YEPD liquid culture medium; the fermentation culture is divided into two stages; in the first stage, the fermentation culture time is 12 hours, and the fermentation culture temperature is 28 DEG C; and in the second stage, the fermentation culture time is 0.5-1 hour, the fermentation culture temperature is 60+/-2 DEG C, and a proteinase is added in the YEPD liquid culture medium, wherein the proteinase is selected from trypsinase, alkaline proteinase and neutral proteinase. The Torulaspora delbrueckii M8 is applicable to the field of microbes.
Description
Technical field
The present invention relates to a Yeasts and improve the method for its fungistatic effect.
Background technology
Yeast is a kind of unicellular microorganism, belongs to the Mycophyta of high microorganism, can survive, belong to facultative anaerobe under aerobic and oxygen-free environment.Yeast is the maximum a kind of microorganism of the direct amount of the mankind.
Synthetic sanitas (or agricultural chemicals) is still China's method the most frequently used to disease control after fruit and vegetable picking, as imazalil, TBZ etc. be everlasting grain plantation and end processing sequences in use.Although this kind of medicament fungistatic effect is remarkable, major injury can be caused to environment to tissue, nontoxic to people and animals, and the research and development with the green preservatives of broad-spectrum antibacterial effect are extremely urgent.
Summary of the invention
The present invention is to solve synthetic sanitas contaminate environment, causing the problems such as major injury to tissue, and provides a kind of broad-spectrum antibacterial yeast and improve the method for its fungistatic effect.
Broad-spectrum antibacterial yeast of the present invention is that Dare is furnished with spore torula (Torulaspora delbrueckii) M8, and be preserved in China typical culture collection center, preserving number is CCTCC NO:M 2013277.
Improve the method for above-mentioned broad-spectrum antibacterial yeast fungistatic effect: Dare is furnished with spore torula (Torulaspora delbrueckii) M8 YEPD solution culture fermentation and cultivates, fermentation culture is divided into two benches, fermentation first stage fermented incubation time is 12h, and fermentation culture temperature is 28 DEG C; Fermentation subordinate phase fermented incubation time is 0.5 ~ 1h, and fermentation culture temperature is 60 ± 2 DEG C, and adds proteolytic enzyme in YEPD liquid nutrient medium, and proteolytic enzyme is selected from trypsinase, Sumizyme MP and neutral protease;
Wherein YEPD liquid nutrient medium is made up of 1g yeast extract paste, 2g peptone, 2g glucose and 100mL distilled water in proportion, and YEPD liquid nutrient medium pH value is 7; Fermentation subordinate phase adds neutral protease YEPD liquid nutrient medium pH value and is adjusted to 7, and fermentation subordinate phase adds trypsinase YEPD liquid nutrient medium pH value and is adjusted to 8, and fermentation subordinate phase adds Sumizyme MP YEPD liquid nutrient medium pH value and is adjusted to 9.
It is cima shape, monolateral budding that Dare is furnished with spore torula (Torulaspora delbrueckii) M8, and thalline diameter is 3.25 μm; Bacterium colony is cima shape, smooth surface, neat in edge, white, opaque, moistening.
Dare is furnished with spore torula (Torulaspora delbrueckii) M8 and all has stable, excellent fungistatic effect to intestinal bacteria, streptococcus aureus, candida albicans, Shigellae, Salmonellas and subtilis.
Yeast existence nutritional needs is simple, and reproduction speed is fast, can not produce the objectionable impurities threatening the mankind and environmental safety, and can promoting digestion after being eaten by human or animal, is therefore natural, safe food preservatives and fungistat.
Dare is furnished with spore torula (Torulaspora delbrueckii) M8 and can survives at drier fruit and vegetable surfaces, and competes nutrition and space with germ, little on the impact of other antiseptic-germicide; And produce antibacterial substance be mostly on fruit, vegetables normal bacteria completion point, to human-body safety.
The antibacterial substance that Dare is furnished with in spore torula (Torulaspora delbrueckii) M8 can add in the drink such as fruit juice, milk as natural antiseptic agent, utilizes its antibacterial quality guaranteed period forming prolongation product
Dare is furnished with can make starter in spore torula (Torulaspora delbrueckii) M8, and its leavened prod can suppress enteron aisle common pathogen.
With cultivate with YEPD solution culture fermentation, fermentation culture temperature is 28 DEG C, fermented incubation time is that the Dare of 13h is furnished with spore torula (Torulaspora delbrueckii) M8 and compares, the inventive method can improve the fungistatic effect (antibacterial circle diameter) that Dare is furnished with spore torula (Torulaspora delbrueckii) M8 and reach more than 2 times, has and significantly improves.
It is that Dare is furnished with spore torula (Torulaspora delbrueckii) that broad-spectrum antibacterial yeast Dare is furnished with spore torula (Torulaspora delbrueckii) M8, belongs to ball spore yeast belong (Torulaspora); Be preserved in China typical culture collection center (CCTCC), preservation address is Wuhan University, and preservation date is on June 23rd, 2013, and preserving number is CCTCC NO:M2013277.
Accompanying drawing explanation
Fig. 1 is the fungistatic effect figure to Shigellae in embodiment 1, and what add in A in Fig. 1 is that to be furnished with what add in spore torula (Torulaspora delbrueckii) M8, B be the Nisin tired as 5IU to Dare.
Fig. 2 is that what add in A in Fig. 2 is that to be furnished with what add in spore torula (Torulaspora delbrueckii) M8, B be the Nisin tired as 5IU to Dare to colibacillary fungistatic effect figure in embodiment 1.
Fig. 3 is the fungistatic effect figure to Candida albicans in embodiment 1, and what add in A in Fig. 3 is that to be furnished with what add in spore torula (Torulaspora delbrueckii) M8, B be the Nisin tired as 5IU to Dare.
Fig. 4 is the fungistatic effect figure to Salmonellas in embodiment 1, and what add in A in Fig. 4 is that to be furnished with what add in spore torula (Torulaspora delbrueckii) M8, B be the Nisin tired as 5IU to Dare.
Fig. 5 is the fungistatic effect figure to subtilis in embodiment 1, and what add in A in Fig. 5 is that to be furnished with what add in spore torula (Torulaspora delbrueckii) M8, B be the Nisin tired as 5IU to Dare.
Fig. 6 is the fungistatic effect figure to streptococcus aureus in embodiment 1, and what add in A in Fig. 6 is that to be furnished with what add in spore torula (Torulaspora delbrueckii) M8, B be the Nisin tired as 5IU to Dare.
Fig. 7 be embodiment 2 1. under subordinate phase different fermentations temperature condition Dare be furnished with spore torula (Torulaspora delbrueckii) M8 to the fungistatic effect figure of Shigellae, in Fig. 7, in A, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase temperature is 28 DEG C, in Fig. 7, in B, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase temperature is 60 DEG C, in Fig. 7, in C, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase temperature is 80 DEG C.
Fig. 8 be embodiment 2 1. under subordinate phase different fermentations temperature condition Dare be furnished with spore torula (Torulaspora delbrueckii) M8 to the fungistatic effect figure of Shigellae, B in Fig. 8
1it is 100 DEG C that middle Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase temperature, C in Fig. 8
1it is 120 DEG C that middle Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase temperature.
Fig. 9 be embodiment 2 1. under subordinate phase different fermentations temperature condition Dare be furnished with spore torula (Torulaspora delbrueckii) M8 to colibacillary fungistatic effect figure, in Fig. 9, in A, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase temperature is 28 DEG C, in Fig. 9, in B, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase temperature is 60 DEG C, in Fig. 9, in C, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase temperature is 80 DEG C, in Fig. 9, in D, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase temperature is 100 DEG C.
Figure 10 be embodiment 2 1. under subordinate phase different fermentations temperature condition Dare be furnished with spore torula (Torulaspora delbrueckii) M8 to colibacillary fungistatic effect figure, in Figure 10, in A, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase temperature is 28 DEG C, in Figure 10, in E, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase temperature is 100 DEG C, in Figure 10, in F, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase temperature is 120 DEG C.
Figure 11 be embodiment 2 2. under subordinate phase different fermentations pH condition Dare be furnished with spore torula (Torulaspora delbrueckii) M8 to the fungistatic effect figure of Shigellae, in Figure 11, in A, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase pH value is 7, in Figure 11, in B, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase pH value is 4, in Figure 11, in C, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase pH value is 6, in Figure 11, in D, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase pH value is 8.
Figure 12 be embodiment 2 2. under subordinate phase different fermentations pH condition Dare be furnished with spore torula (Torulaspora delbrueckii) M8 to the fungistatic effect figure of Shigellae, in Figure 11, in A, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase pH value is 7, in Figure 11, in E, Dare is furnished with ferment subordinate phase pH value of spore torula (Torulaspora delbrueckii) M8 be that in 10, Figure 11, in F, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 subordinate phase pH value of fermenting is 12.
Figure 13 be embodiment 2 2. under subordinate phase different fermentations pH condition Dare be furnished with spore torula (Torulaspora delbrueckii) M8 to colibacillary fungistatic effect figure, in Figure 13, in A, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase pH value is 7, in Figure 13, in B, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase pH value is 4, in Figure 13, in C, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase pH value is 6, in Figure 13, in D, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase pH value is 8.
Figure 14 be embodiment 2 2. under subordinate phase different fermentations pH condition Dare be furnished with spore torula (Torulaspora delbrueckii) M8 to colibacillary fungistatic effect figure, in Figure 14, in A, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermentation subordinate phase pH value is 7, in Figure 14, in E, Dare is furnished with ferment subordinate phase pH value of spore torula (Torulaspora delbrueckii) M8 be that in 10, Figure 14, in F, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 subordinate phase pH value of fermenting is 12.
Figure 15 be embodiment 2 3. in subordinate phase add different proteolytic enzyme condition under Dare be furnished with spore torula (Torulaspora delbrueckii) M8 to the fungistatic effect figure of Shigellae, in Figure 15 in A Dare be furnished with spore torula (Torulaspora delbrueckii) M8 ferment subordinate phase do not add proteolytic enzyme, in Figure 15 in B Dare be furnished with spore torula (Torulaspora delbrueckii) M8 ferment subordinate phase add trypsinase, in Figure 15 in C Dare be furnished with spore torula (Torulaspora delbrueckii) M8 ferment subordinate phase add neutral protease, in Figure 15 in D Dare be furnished with spore torula (Torulaspora delbrueckii) M8 ferment subordinate phase add Sumizyme MP.
Figure 16 be embodiment 2 3. in subordinate phase add different proteolytic enzyme condition under Dare be furnished with spore torula (Torulaspora delbrueckii) M8 to colibacillary fungistatic effect figure, in Figure 16 in A Dare be furnished with spore torula (Torulaspora delbrueckii) M8 ferment subordinate phase do not add proteolytic enzyme, in Figure 16 in B Dare be furnished with spore torula (Torulaspora delbrueckii) M8 ferment subordinate phase add trypsinase, in Figure 16 in C Dare be furnished with spore torula (Torulaspora delbrueckii) M8 ferment subordinate phase add neutral protease, in Figure 16 in D Dare be furnished with spore torula (Torulaspora delbrueckii) M8 ferment subordinate phase add Sumizyme MP.
Figure 17 is the phylogeny tree graph that Dare is furnished with that spore torula (Torulaspora delbrueckii) M8 and close bacterial strain carry out sequence analysis structure.
Embodiment
Technical solution of the present invention is not limited to following cited embodiment, also comprises the arbitrary combination between each embodiment.
Embodiment one: present embodiment broad-spectrum antibacterial yeast is that Dare is furnished with spore torula (Torulaspora delbrueckii) M8, and be preserved in China typical culture collection center, preserving number is CCTCC NO:M 2013277.
Dare is furnished with spore torula (Torulaspora delbrueckii) M8 and gathers from Tibet, is separated and obtains from China's traditional zymotic milk-product tibet koumiss.Tibet koumiss is placed in the skimming milk Secondary Culture of 10%, after activation, its fermented liquid is applied in the YEPD solid medium containing paraxin separation of ruling and obtains Dare and be furnished with spore torula (Torulaspora delbrueckii) M8.
The 26s rDNA D1/D2 district bases longs that Dare is furnished with spore torula (Torulaspora delbrueckii) M8 is 566bp, base sequence is as shown in SEQ ID NO:1,18S rDNA length is 1192bp, and base sequence is as shown in SEQ ID NO:2; Through 26S rDNA D1/D2 region sequence and 18S rDNA sequence alignment, and by microbial morphology and physiological and biochemical index qualification, confirm that broad-spectrum antibacterial yeast M8 is that ball spore yeast belong (Torulaspora) Dare is furnished with spore torula (Torulaspora delbrueckii), the most close with Torulaspora delbrueckii CBS 1146 homology.
The biochemical results of Torulaspora delbrueckii CBS 1146 is as shown in table 1, and Torulaspora delbrueckii CBS 1146 bacterium colony is yellow or purple.
Table 1
The biochemical results that Dare is furnished with spore torula (Torulaspora delbrueckii) M8 is as shown in table 2, and bacterium colony is white.
Table 2
Compare according to biochemical test result and colonial morphology etc., can determine that Dare is furnished with spore torula (Torulaspora delbrueckii) M8 is that a strain Dare is furnished with spore torula new strains.
Dare is furnished with spore torula (Torulaspora delbrueckii) M8 can suppress intestinal bacteria, streptococcus aureus, candida albicans, Shigellae, Salmonellas and subtilis, can be used for food fermentation, suppress the growth of harmful bacteria, reduce pathogenic bacterium to the harm of human body.
Dare is furnished with spore torula (Torulaspora delbrueckii) M8 and can cultivates with YEPD solid medium, preserve, YEPD solid medium is made up of 1g yeast extract paste, 2g peptone, 2g glucose, 2g agar and 100mL distilled water in proportion, and to reconcile pH value be 7.The culture temperature that Dare is furnished with spore torula (Torulaspora delbrueckii) M8 is 28 ~ 30 DEG C.
Dare be furnished with spore torula (Torulaspora delbrueckii) M8 gather from traditional cultured milk prod tibet koumiss, to people and animals and environment nontoxic.
Embodiment two: present embodiment improves the method for above-mentioned broad-spectrum antibacterial yeast fungistatic effect: Dare is furnished with spore torula (Torulaspora delbrueckii) M8 YEPD solution culture fermentation and cultivates, fermentation culture is divided into two benches, fermentation first stage fermented incubation time is 12h, and fermentation culture temperature is 28 DEG C; Fermentation subordinate phase fermented incubation time is 0.5 ~ 1h, and fermentation culture temperature is 60 ± 2 DEG C, and adds proteolytic enzyme in YEPD liquid nutrient medium, and proteolytic enzyme is selected from trypsinase, Sumizyme MP and neutral protease;
Wherein YEPD liquid nutrient medium is made up of 1g yeast extract paste, 2g peptone, 2g glucose and 100mL distilled water in proportion, and YEPD liquid nutrient medium pH value is 7; Fermentation subordinate phase adds neutral protease YEPD liquid nutrient medium pH value and is adjusted to 7, and fermentation subordinate phase adds trypsinase YEPD liquid nutrient medium pH value and is adjusted to 8, and fermentation subordinate phase adds Sumizyme MP YEPD liquid nutrient medium pH value and is adjusted to 9.
Embodiment three: the difference of present embodiment and embodiment two is: add 1.5mg proteolytic enzyme in often liter of YEPD liquid nutrient medium, tryptic enzyme activity is 250U/mg, the enzyme activity of Sumizyme MP is 200U/mg, the enzyme activity of neutral protease is 60U/mg.Other steps and parameter identical with embodiment two.
Embodiment 1 bacteriostatic experiment:
Making bacteriostatic experiment is dull and stereotyped: in sterilizing flat board, pour 8mL element agar into, put Oxford cup after to be solidified on agar; 4ml physiological saline is added respectively in the slant medium containing intestinal bacteria, streptococcus aureus, candida albicans, Shigellae, Salmonellas and subtilis, then shake, therefrom get 0.5ml again and inject 4.5ml stroke-physiological saline solution, abundant concussion, repeat above-mentioned gradient dilution 3 times again, preparation 10
-3bacteria suspension; Add in 25ml nutritional medium by vortex oscillator by getting 1ml after bacteria suspension concussion evenly, mixing is poured in above-mentioned stand-by agar plate, Deng extracting Oxford cup after agar solidification, in left hole, add the concentrated broth that 100 μ l Dares are furnished with spore torula (Torulaspora delbrueckii) M8 (in left hole, add 100 μ l Nisin solution, complete Nisin contrast), namely make bacteriostatic experiment flat board;
Wherein, described plain agar is added in 200ml distilled water by 4g agar and makes;
Described nutritional medium 1000mL is made up of the distilled water of 3 ~ 5g extractum carnis, 10g peptone, 5g sodium-chlor, 7.5g agar and surplus, and reconciling pH value is sterilizing 20min under 7.0,121 DEG C of conditions;
Dare is furnished with the concentrated broth of spore torula (Torulaspora delbrueckii) M8: Dare is furnished with spore torula (Torulaspora delbrueckii) M8 YEPD solution culture fermentation and cultivates, fermentation culture is divided into two benches, fermentation first stage fermented incubation time is 12h, and fermentation culture temperature is 28 DEG C; Fermentation subordinate phase fermented incubation time is 1h, fermentation culture Medium's PH Value is 7, and fermentation culture temperature is 60 DEG C, and adds neutral protease in YEPD liquid nutrient medium, often liter of YEPD liquid nutrient medium adds 1.5g neutral protease, and the enzyme of neutral protease is lived as 60U/mg; Then Dare is furnished with spore torula (Torulaspora delbrueckii) M8 fermented liquid centrifugal 10min under 4000r/min condition, supernatant liquor is placed in Rotary Evaporators vacuum tightness 660mmHg, is concentrated into 1/16 of original volume at 55 DEG C, namely obtain the concentrated broth that Dare is furnished with spore torula (Torulaspora delbrueckii) M8;
Wherein, described Nisin solution is that 110mgNisin adds in 10mL water, and after dissolving, constant volume is to 100mL, namely obtains Nisin solution.
The constant incubator that bacteriostatic experiment flat board is placed in 37 DEG C is cultivated 12h, and then observe, fungistatic effect is as shown in Fig. 1 ~ Fig. 6, and it is as shown in table 3 that Dare is furnished with spore torula (Torulaspora delbrueckii) M8 inhibition zone data.
Table 3
Experimental result shows that the fungistatic effect that Dare is furnished with spore torula (Torulaspora delbrueckii) M8 is: be 24.30mm to the antibacterial circle diameter mean value of Shigellae, and inhibition zone is very clear, transparent; To colibacillary antibacterial circle diameter mean value 23.72mm, inhibition zone is clear, transparent; Be 23.08mm to the antibacterial circle diameter mean value of candida albicans, inhibition zone is clear, transparent; Be 18.53mm to the antibacterial circle diameter mean value of Salmonellas, inhibition zone is relatively clear, transparent; Be 16.68mm to the antibacterial circle diameter mean value of subtilis, inhibition zone is clear, transparent; Be 16.66mm to the antibacterial circle diameter mean value of streptococcus aureus, inhibition zone is generally clear, transparent.Dare is furnished with spore torula (Torulaspora delbrueckii) M8 all has good fungistatic effect to intestinal bacteria, streptococcus aureus, candida albicans, Shigellae, Salmonellas and subtilis, illustrates that Dare is furnished with spore torula (Torulaspora delbrueckii) M8 and has broad-spectrum antibacterial.
Embodiment 2 improves fungistatic effect experiment:
1. Dare is furnished with the cultivation of spore torula (Torulaspora delbrueckii) M8 YEPD solution culture fermentation, and fermentation culture is divided into two benches, and fermentation first stage fermented incubation time is 12h, and fermentation culture temperature is 28 DEG C; Fermentation subordinate phase respectively in 120 DEG C, 100 DEG C, 80 DEG C, 60 DEG C and 28 DEG C of environment fermented incubation time be 0.5h, and add neutral protease (enzyme activity of neutral protease is 60U/mg);
Wherein YEPD liquid nutrient medium is made up of 1g yeast extract paste, 2g peptone, 2g glucose and 100mL distilled water in proportion, and YEPD liquid nutrient medium pH value is 7.
In sterilizing flat board, pour 8mL element agar into, after to be solidified, on agar, put Oxford cup; Add 4ml physiological saline respectively to containing in Shigellae and colibacillary slant medium, then shake, more therefrom get 0.5ml injection 4.5ml stroke-physiological saline solution, fully shake, then repeat above-mentioned gradient dilution 3 times, preparation 10
-3bacteria suspension; Add in 25ml nutritional medium by vortex oscillator by getting 1ml after bacteria suspension concussion evenly, mixing is poured in above-mentioned stand-by agar plate, Deng extracting Oxford cup after agar solidification, in left hole, add the concentrated broth that above-mentioned 100 μ l Dares are furnished with spore torula (Torulaspora delbrueckii) M8 respectively;
Wherein, described plain agar is added in 200ml distilled water by 4g agar and makes;
Described nutritional medium 1000mL is made up of the distilled water of 3 ~ 5g extractum carnis, 10g peptone, 5g sodium-chlor, 7.5g agar and surplus, and reconciling pH value is sterilizing 20min under 7.0,121 DEG C of conditions;
Dare is furnished with the concentrated broth of spore torula (Torulaspora delbrueckii) M8: the Dare cultivated under different condition is furnished with spore torula (Torulaspora delbrueckii) M8 fermented liquid centrifugal 10min under 4000r/min condition, supernatant liquor is placed in Rotary Evaporators vacuum tightness 660mmHg, is concentrated into 1/16 of original volume at 55 DEG C, namely obtain the concentrated broth that Dare is furnished with spore torula (Torulaspora delbrueckii) M8.
The constant incubator above-mentioned assay plate being placed in 37 DEG C cultivates 12h, then observe, under fermentation subordinate phase different fermentations temperature condition, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 to the fungistatic effect of Shigellae as shown in Figure 7 and Figure 8, to colibacillary fungistatic effect as shown in Figure 9 and Figure 10.
Experimental result: fermentation subordinate phase temperature is that to be furnished with spore torula (Torulaspora delbrueckii) M8 be 19.031mm to the antibacterial circle diameter mean value that Shigellae produces to the Dare of 28 DEG C, fermentation subordinate phase temperature is that to be furnished with spore torula (Torulaspora delbrueckii) M8 be 20.158mm to the antibacterial circle diameter mean value that Shigellae produces to the Dare of 60 DEG C, fermentation subordinate phase temperature is that to be furnished with spore torula (Torulaspora delbrueckii) M8 be 15.405mm to the antibacterial circle diameter mean value that Shigellae produces to the Dare of 80 DEG C, fermentation subordinate phase temperature is that to be furnished with spore torula (Torulaspora delbrueckii) M8 be 8.941mm to the antibacterial circle diameter mean value that Shigellae produces to the Dare of 100 DEG C, fermentation subordinate phase temperature is that to be furnished with spore torula (Torulaspora delbrueckii) M8 be 0mm to the antibacterial circle diameter mean value that Shigellae produces to the Dare of 120 DEG C.
Fermentation subordinate phase temperature is that the Dare of 28 DEG C is furnished with spore torula (Torulaspora delbrueckii) M8 and is respectively 19.363mm and 18.762mm to the antibacterial circle diameter that intestinal bacteria produce, fermentation subordinate phase temperature is that to be furnished with spore torula (Torulaspora delbrueckii) M8 be 20.957mm to the antibacterial circle diameter mean value that intestinal bacteria produce to the Dare of 60 DEG C, fermentation subordinate phase temperature is that to be furnished with spore torula (Torulaspora delbrueckii) M8 be 10.168mm to the antibacterial circle diameter mean value that intestinal bacteria produce to the Dare of 80 DEG C, fermentation subordinate phase temperature is that the Dare of 100 DEG C is furnished with spore torula (Torulaspora delbrueckii) M8 and is respectively 10.547mm and 7.516mm to the antibacterial circle diameter that intestinal bacteria produce, fermentation subordinate phase temperature is that to be furnished with spore torula (Torulaspora delbrueckii) M8 be 0mm to the antibacterial circle diameter mean value that intestinal bacteria produce to the Dare of 120 DEG C.
This test fermentation subordinate phase temperature is that the fungistatic effect that 60 DEG C of Dares are furnished with spore torula (Torulaspora delbrueckii) M8 improves the most remarkable.
2. Dare is furnished with the cultivation of spore torula (Torulaspora delbrueckii) M8 YEPD solution culture fermentation, and fermentation culture is divided into two benches, and fermentation first stage fermented incubation time is 12h, and fermentation culture temperature is 28 DEG C; Fermentation subordinate phase fermented incubation time is 1h, and fermentation culture temperature is 28 DEG C;
Wherein YEPD liquid nutrient medium is made up of 1g yeast extract paste, 2g peptone, 2g glucose and 100mL distilled water in proportion, fermentation first stage YEPD liquid nutrient medium pH value is 5.5, fermentation subordinate phase YEPD liquid nutrient medium regulates fermented liquid pH value to be pH to be 12,10,8,7,6,4, to be adjusted to neutrality again before experiment respectively.
In sterilizing flat board, pour 8mL element agar into, after to be solidified, on agar, put Oxford cup; Add 4ml physiological saline respectively to containing in Shigellae and colibacillary slant medium, then shake, more therefrom get 0.5ml injection 4.5ml stroke-physiological saline solution, fully shake, then repeat above-mentioned gradient dilution 3 times, preparation 10
-3bacteria suspension; Add in 25ml nutritional medium by vortex oscillator by getting 1ml after bacteria suspension concussion evenly, mixing is poured in above-mentioned stand-by agar plate, Deng extracting Oxford cup after agar solidification, in left hole, add the concentrated broth that above-mentioned 100 μ l Dares are furnished with spore torula (Torulaspora delbrueckii) M8 respectively;
Wherein, described plain agar is added in 200ml distilled water by 4g agar and makes;
Described nutritional medium 1000mL is made up of the distilled water of 3 ~ 5g extractum carnis, 10g peptone, 5g sodium-chlor, 7.5g agar and surplus, and reconciling pH value is sterilizing 20min under 7.0,121 DEG C of conditions;
Dare is furnished with the concentrated broth of spore torula (Torulaspora delbrueckii) M8: the Dare cultivated under different condition is furnished with spore torula (Torulaspora delbrueckii) M8 fermented liquid centrifugal 10min under 4000r/min condition, supernatant liquor is placed in Rotary Evaporators vacuum tightness 660mmHg, is concentrated into 1/16 of original volume at 55 DEG C, namely obtain the concentrated broth that Dare is furnished with spore torula (Torulaspora delbrueckii) M8.
The constant incubator above-mentioned assay plate being placed in 37 DEG C cultivates 12h, then observe, under fermentation subordinate phase different fermentations pH condition, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 to the fungistatic effect of Shigellae as is illustrated by figs. 11 and 12, to colibacillary fungistatic effect as shown in Figure 13 and Figure 14.
Experimental result: fermentation subordinate phase pH value be 4 Dare to be furnished with spore torula (Torulaspora delbrueckii) M8 be 11.362mm to the antibacterial circle diameter mean value that Shigellae produces, fermentation subordinate phase pH value be 6 Dare to be furnished with spore torula (Torulaspora delbrueckii) M8 be 12.112mm to the antibacterial circle diameter mean value that Shigellae produces, fermentation subordinate phase pH value be 7 Dare be furnished with spore torula (Torulaspora delbrueckii) M8 12.136mm and 14.175mm be respectively to the antibacterial circle diameter that Shigellae produces, fermentation subordinate phase pH value be 8 Dare to be furnished with spore torula (Torulaspora delbrueckii) M8 be 9.158mm to the antibacterial circle diameter mean value that Shigellae produces, fermentation subordinate phase pH value be 10 Dare to be furnished with spore torula (Torulaspora delbrueckii) M8 be 7.693mm to the antibacterial circle diameter mean value that Shigellae produces, fermentation subordinate phase pH value be 12 Dare to be furnished with spore torula (Torulaspora delbrueckii) M8 be 0.021mm to the antibacterial circle diameter mean value that Shigellae produces.
Fermentation subordinate phase pH value be 4 Dare to be furnished with spore torula (Torulaspora delbrueckii) M8 be 10.261mm to the antibacterial circle diameter mean value that intestinal bacteria produce, fermentation subordinate phase pH value be 6 Dare to be furnished with spore torula (Torulaspora delbrueckii) M8 be 14.245mm to the antibacterial circle diameter mean value that intestinal bacteria produce, fermentation subordinate phase pH value be 7 Dare be furnished with spore torula (Torulaspora delbrueckii) M8 14.367mm and 15.863mm be respectively to the antibacterial circle diameter that intestinal bacteria produce, fermentation subordinate phase pH value be 8 Dare to be furnished with spore torula (Torulaspora delbrueckii) M8 be 13.638mm to the antibacterial circle diameter mean value that intestinal bacteria produce, fermentation subordinate phase pH value be 10 Dare to be furnished with spore torula (Torulaspora delbrueckii) M8 be 5.053mm to the antibacterial circle diameter mean value that intestinal bacteria produce, fermentation subordinate phase pH value be 12 Dare to be furnished with spore torula (Torulaspora delbrueckii) M8 be 0mm to the antibacterial circle diameter mean value that intestinal bacteria produce.
This test fermentation subordinate phase pH value is that the fungistatic effect that 7 Dares are furnished with spore torula (Torulaspora delbrueckii) M8 improves the most remarkable.
3. Dare is furnished with the cultivation of spore torula (Torulaspora delbrueckii) M8 YEPD solution culture fermentation, and fermentation culture is divided into two benches, and fermentation first stage fermented incubation time is 12h, and fermentation culture temperature is 28 DEG C; Fermentation subordinate phase fermented incubation time is 1h, and adds proteolytic enzyme (what do not add proteolytic enzyme is set to contrast) in the ratio of often liter of YEPD liquid nutrient medium 1.5g proteolytic enzyme in fermentation subordinate phase;
Wherein YEPD liquid nutrient medium is made up of 1g yeast extract paste, 2g peptone, 2g glucose and 100mL distilled water in proportion, and fermentation first stage YEPD liquid nutrient medium pH value is 7;
First group of fermentation subordinate phase adds trypsinase, and fermentation subordinate phase leavening temperature is 37 DEG C, and fermentation subordinate phase YEPD liquid nutrient medium pH value is 8, and often liter of YEPD liquid nutrient medium adds 1.5mg trypsinase, and tryptic enzyme is lived as 250U/mg;
Second group of fermentation subordinate phase adds neutral protease, and fermentation subordinate phase leavening temperature is 40 DEG C, and fermentation subordinate phase YEPD liquid nutrient medium pH value is 7, and often liter of YEPD liquid nutrient medium adds 1.5mg neutral protease, and the enzyme of neutral protease is lived as 60U/mg;
3rd group of fermentation subordinate phase adds Sumizyme MP, and fermentation subordinate phase leavening temperature is 40 DEG C, and fermentation subordinate phase YEPD liquid nutrient medium pH value is 9, and often liter of YEPD liquid nutrient medium adds 1.5mg Sumizyme MP, and the enzyme of Sumizyme MP is lived as 200U/mg;
4th group is control group, does not add proteolytic enzyme, and fermentation culture temperature is 28 DEG C, and fermented incubation time is 1h.
In sterilizing flat board, pour 8mL element agar into, after to be solidified, on agar, put Oxford cup; Add 4ml physiological saline respectively to containing in Shigellae and colibacillary slant medium, then shake, more therefrom get 0.5ml injection 4.5ml stroke-physiological saline solution, fully shake, then repeat above-mentioned gradient dilution 3 times, preparation 10
-3bacteria suspension; Add in 25ml nutritional medium by vortex oscillator by getting 1ml after bacteria suspension concussion evenly, mixing is poured in above-mentioned stand-by agar plate, Deng extracting Oxford cup after agar solidification, in left hole, add the concentrated broth that above-mentioned 100 μ l Dares are furnished with spore torula (Torulaspora delbrueckii) M8 respectively;
Wherein, described plain agar is added in 200ml distilled water by 4g agar and makes;
Described nutritional medium 1000mL is made up of the distilled water of 3 ~ 5g extractum carnis, 10g peptone, 5g sodium-chlor, 7.5g agar and surplus, and reconciling pH value is sterilizing 20min under 7.0,121 DEG C of conditions;
Dare is furnished with the concentrated broth of spore torula (Torulaspora delbrueckii) M8: the Dare cultivated under different condition is furnished with spore torula (Torulaspora delbrueckii) M8 fermented liquid centrifugal 10min under 4000r/min condition, supernatant liquor is placed in Rotary Evaporators vacuum tightness 660mmHg, is concentrated into 1/16 of original volume at 55 DEG C, namely obtain the concentrated broth that Dare is furnished with spore torula (Torulaspora delbrueckii) M8.
The constant incubator above-mentioned assay plate being placed in 37 DEG C cultivates 12h, then observe, under fermentation subordinate phase adds different proteolytic enzyme condition, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 to the fungistatic effect of Shigellae as shown in figure 15, to colibacillary fungistatic effect as shown in figure 16.
Experimental result: it is 10.853mm to the antibacterial circle diameter mean value that Shigellae produces that the Dare that fermentation subordinate phase does not add proteolytic enzyme is furnished with spore torula (Torulaspora delbrueckii) M8, fermentation subordinate phase adds tryptic Dare, and to be furnished with spore torula (Torulaspora delbrueckii) M8 be 15.913mm to the antibacterial circle diameter mean value that Shigellae produces, it is 14.139mm to the antibacterial circle diameter mean value that Shigellae produces that the Dare that fermentation subordinate phase adds neutral protease is furnished with spore torula (Torulaspora delbrueckii) M8, it is 13.227mm to the antibacterial circle diameter mean value that Shigellae produces that the Dare that fermentation subordinate phase adds Sumizyme MP is furnished with spore torula (Torulaspora delbrueckii) M8.
It is 9.471mm to the antibacterial circle diameter mean value that intestinal bacteria produce that the Dare that fermentation subordinate phase does not add proteolytic enzyme is furnished with spore torula (Torulaspora delbrueckii) M8, fermentation subordinate phase adds tryptic Dare, and to be furnished with spore torula (Torulaspora delbrueckii) M8 be 16.974mm to the antibacterial circle diameter mean value that intestinal bacteria produce, it is 15.843mm to the antibacterial circle diameter mean value that intestinal bacteria produce that the Dare that fermentation subordinate phase adds neutral protease is furnished with spore torula (Torulaspora delbrueckii) M8, it is 16.159mm to the antibacterial circle diameter mean value that intestinal bacteria produce that the Dare that fermentation subordinate phase adds Sumizyme MP is furnished with spore torula (Torulaspora delbrueckii) M8.
4. Dare is furnished with the cultivation of spore torula (Torulaspora delbrueckii) M8 YEPD solution culture fermentation, and fermentation culture is divided into two benches, and fermentation first stage fermented incubation time is 12h, and fermentation culture temperature is 28 DEG C; Fermentation subordinate phase fermented incubation time is 1h, and adds proteolytic enzyme (what do not add proteolytic enzyme is set to contrast) in the ratio of often liter of YEPD liquid nutrient medium 1.5g proteolytic enzyme in fermentation subordinate phase;
Wherein YEPD liquid nutrient medium is made up of 1g yeast extract paste, 2g peptone, 2g glucose and 100mL distilled water in proportion, and fermentation first stage YEPD liquid nutrient medium pH value is 7;
First group of fermentation subordinate phase leavening temperature is 40 DEG C, and first regulate YEPD liquid nutrient medium pH value to be 9 add Sumizyme MP 1mg, the enzyme of Sumizyme MP is lived as 200U/mg, and fermented incubation time is 0.5h; Then regulate YEPD liquid nutrient medium pH value to be 7 add neutral protease 0.5mg, the enzyme of neutral protease is lived as 60U/mg, and continuation fermented incubation time is 0.5h;
Second group first regulates YEPD liquid nutrient medium pH value to be 8 add trypsinase 0.5mg, and tryptic enzyme is lived as 250U/mg, cultivates 0.5h at 37 DEG C of condition bottom fermentations; Then regulate YEPD liquid nutrient medium pH value to be 9 add Sumizyme MP 1mg, the enzyme of Sumizyme MP is lived as 200U/mg, and under 40 DEG C of conditions, continue fermented incubation time is 0.5h;
In sterilizing flat board, pour 8mL element agar into, after to be solidified, on agar, put Oxford cup; Add 4ml physiological saline respectively to containing in Shigellae and colibacillary slant medium, then shake, more therefrom get 0.5ml injection 4.5ml stroke-physiological saline solution, fully shake, then repeat above-mentioned gradient dilution 3 times, preparation 10
-3bacteria suspension; Add in 25ml nutritional medium by vortex oscillator by getting 1ml after bacteria suspension concussion evenly, mixing is poured in above-mentioned stand-by agar plate, Deng extracting Oxford cup after agar solidification, in left hole, add the concentrated broth that above-mentioned 100 μ l Dares are furnished with spore torula (Torulaspora delbrueckii) M8 respectively;
Wherein, described plain agar is added in 200ml distilled water by 4g agar and makes;
Described nutritional medium 1000mL is made up of the distilled water of 3 ~ 5g extractum carnis, 10g peptone, 5g sodium-chlor, 7.5g agar and surplus, and reconciling pH value is sterilizing 20min under 7.0,121 DEG C of conditions;
Dare is furnished with the concentrated broth of spore torula (Torulaspora delbrueckii) M8: the Dare cultivated under different condition is furnished with spore torula (Torulaspora delbrueckii) M8 fermented liquid centrifugal 10min under 4000r/min condition, supernatant liquor is placed in Rotary Evaporators vacuum tightness 660mmHg, is concentrated into 1/16 of original volume at 55 DEG C, namely obtain the concentrated broth that Dare is furnished with spore torula (Torulaspora delbrueckii) M8.
The constant incubator above-mentioned assay plate being placed in 37 DEG C cultivates 12h, experimental result: first group of antibacterial circle diameter mean value produced Shigellae is 14.002mm, and the antibacterial circle diameter mean value produced intestinal bacteria is 16.027mm; Second group of antibacterial circle diameter mean value produced Shigellae is 14.522mm, and the antibacterial circle diameter mean value produced intestinal bacteria is 16.536mm.
The fungistatic effect that the Dare that this test fermentation subordinate phase adds proteolytic enzyme is furnished with spore torula (Torulaspora delbrueckii) M8 is significantly increased.
Claims (3)
1. a broad-spectrum antibacterial yeast, it is furnished with spore torula (Torulaspora delbrueckii) M8 for Dare, and be preserved in China typical culture collection center, preserving number is CCTCC NO:M 2013277.
2. improve the method for broad-spectrum antibacterial yeast fungistatic effect described in claim 1, Dare is furnished with spore torula (Torulaspora delbrueckii) M8 YEPD solution culture fermentation and cultivates, it is characterized in that fermentation culture is divided into two benches, fermentation first stage fermented incubation time is 12h, and fermentation culture temperature is 28 DEG C; Fermentation subordinate phase fermented incubation time is 0.5 ~ 1h, and fermentation culture temperature is 60 ± 2 DEG C, and adds proteolytic enzyme in YEPD liquid nutrient medium, and proteolytic enzyme is selected from trypsinase, Sumizyme MP and neutral protease;
Wherein YEPD liquid nutrient medium is made up of 1g yeast extract paste, 2g peptone, 2g glucose and 100mL distilled water in proportion, and YEPD liquid nutrient medium pH value is 7; Fermentation subordinate phase adds neutral protease YEPD liquid nutrient medium pH value and is adjusted to 7, and fermentation subordinate phase adds trypsinase YEPD liquid nutrient medium pH value and is adjusted to 8, and fermentation subordinate phase adds Sumizyme MP YEPD liquid nutrient medium pH value and is adjusted to 9.
3. the method for raising broad-spectrum antibacterial yeast fungistatic effect according to claim 2, described fermentation subordinate phase often rises in YEPD liquid nutrient medium and adds 1.5mg proteolytic enzyme, and tryptic enzyme activity is 250U/mg, the enzyme activity of Sumizyme MP is 200U/mg, the enzyme activity of neutral protease is 60U/mg.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410449239.7A CN104263662B (en) | 2014-04-01 | 2014-09-05 | Broad-spectrum antibacterial microzyme and method for enhancing antibacterial effect thereof |
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410128540.8 | 2014-04-01 | ||
| CN2014101285408 | 2014-04-01 | ||
| CN201410128540 | 2014-04-01 | ||
| CN201410449239.7A CN104263662B (en) | 2014-04-01 | 2014-09-05 | Broad-spectrum antibacterial microzyme and method for enhancing antibacterial effect thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104263662A true CN104263662A (en) | 2015-01-07 |
| CN104263662B CN104263662B (en) | 2017-02-15 |
Family
ID=52155246
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410449239.7A Active CN104263662B (en) | 2014-04-01 | 2014-09-05 | Broad-spectrum antibacterial microzyme and method for enhancing antibacterial effect thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104263662B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105062903A (en) * | 2015-09-02 | 2015-11-18 | 河南工业大学 | Low-temperature-resistant high-sugar-yield Torulaspora delbrueckii strain and application thereof |
| CN109042855A (en) * | 2018-08-30 | 2018-12-21 | 河南科技大学 | A kind of application of fruit and vegetable fresh-keeping agent and trypsase in terms of promoting candidiasis Biocontrol Effect |
| CN109337841A (en) * | 2017-12-27 | 2019-02-15 | 山东农业大学 | One plant of bacillus subtilis BYS2 with Efficient antibacterial performance |
| CN111172085A (en) * | 2020-03-15 | 2020-05-19 | 青岛宝创生物科技有限公司 | Application of bacillus subtilis in production of substances inhibiting fungi |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008105516A1 (en) * | 2007-02-28 | 2008-09-04 | Japan Tobacco Inc. | Antifungal substance |
-
2014
- 2014-09-05 CN CN201410449239.7A patent/CN104263662B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008105516A1 (en) * | 2007-02-28 | 2008-09-04 | Japan Tobacco Inc. | Antifungal substance |
Non-Patent Citations (2)
| Title |
|---|
| M. PSANI，P. KOTZEKIDOU: "Technological characteristics of yeast strains and their potential as starter adjuncts in Greek-style black olive fermentation", 《WORLD J MICROBIOL BIOTECHNOL》 * |
| T. SILVA,ET AL: "Characterization of yeasts from Portuguese brined olives, with a focus on their potentially probiotic behavior", 《FOOD SCIENCE AND TECHNOLOGY》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105062903A (en) * | 2015-09-02 | 2015-11-18 | 河南工业大学 | Low-temperature-resistant high-sugar-yield Torulaspora delbrueckii strain and application thereof |
| CN105062903B (en) * | 2015-09-02 | 2018-02-02 | 河南工业大学 | A kind of low temperature resistant high yield sugar torulaspora delbrueckii bacterium and its application |
| CN109337841A (en) * | 2017-12-27 | 2019-02-15 | 山东农业大学 | One plant of bacillus subtilis BYS2 with Efficient antibacterial performance |
| CN109042855A (en) * | 2018-08-30 | 2018-12-21 | 河南科技大学 | A kind of application of fruit and vegetable fresh-keeping agent and trypsase in terms of promoting candidiasis Biocontrol Effect |
| CN111172085A (en) * | 2020-03-15 | 2020-05-19 | 青岛宝创生物科技有限公司 | Application of bacillus subtilis in production of substances inhibiting fungi |
| CN111172085B (en) * | 2020-03-15 | 2021-04-02 | 青岛宝创生物科技有限公司 | Application of bacillus subtilis in production of substances inhibiting fungi |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104263662B (en) | 2017-02-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107245457A (en) | A kind of extracting method and application of dendrobium candidum endogenetic fungal bacterial strain and its exocellular polysaccharide of generation and the exocellular polysaccharide | |
| CN103190538B (en) | Aflatoxin B1 degradation agent and application thereof | |
| CN105670958B (en) | One plant of low temperature biocontrol bacterial strain and its application | |
| CN103865837A (en) | Streptomyces having antagonistic effect on penicillium viridicatum | |
| CN103589655A (en) | Preparation method for Bacillus amyloliquefaciens HRH 317 and antibacterial substances thereof | |
| CN101503708B (en) | Cultivation fermentation method of Bacillus coagulans antimycotics active substance | |
| CN104031960A (en) | Extracting method and application of extracellular antimicrobial lipopeptides of bacillus amyloliquefaciens | |
| CN101886047B (en) | Sophorolipid fruit preservative and use thereof in fruit preservation | |
| CN104263662B (en) | Broad-spectrum antibacterial microzyme and method for enhancing antibacterial effect thereof | |
| CN101935628B (en) | Bacillus subtilis and application thereof | |
| CN107974421A (en) | A kind of lactobacillus acidophilus and its screening technique and application, a kind of microbial inoculum | |
| CN105567624A (en) | Synergist for lactococcus lactis subsp. lactis capable of generating nisin through fermentation and use method of synergist | |
| CN105613707A (en) | Bacillus subtilis biological preservative and application thereof to preservation of large yellow croakers | |
| CN103396956A (en) | Saccharomyces cerevisiae, screening and culture methods thereof and bean meal fermentation method thereof | |
| Banerjee et al. | Psychrophilic microorganisms: habitats and exploitation potentials | |
| CN104988082A (en) | Debaryomyces hansenii strain and applications thereof | |
| CN105494445A (en) | Preparation for preventing and controlling pepper blight and application | |
| CN104531543B (en) | Grey green soy bean endogenetic fungus TPL25 and its application in controlling plant diseases | |
| CN104745510B (en) | A kind of lactobacillus fermenti and its application in fungal diseases of plants preventing and treating | |
| CN102321540A (en) | Preparation method of lactic acid Amphibacillus fermentation liquor, and preparation method of lactic acid Amphibacillus fermentation liquor composite antiseptic by using fermentation liquor thereof | |
| CN105018354B (en) | One plant of pick up the ears trichoderma and its application | |
| CN105002100A (en) | Antagonistic yeast, dry powder of antagonistic yeast and use of dry powder of antagonistic yeast | |
| CN104152362B (en) | A kind of broad-spectrum antibacterial saccharomycete | |
| CN109679881A (en) | A kind of general bacterium of pineapple and its microbial inoculum and application | |
| CN102433285B (en) | Bacterium paenibacillus polymyma SHL-1 and application thereof in preventing and controlling stalk rot of cymbidium sinense |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20211014 Address after: Room 1025, building 3, No. 3688, Tingwei Road, Caojing Town, Jinshan District, Shanghai 201500 Patentee after: Shanghai aiang Biotechnology Co.,Ltd. Address before: 150080 No. 74, Xuefu Road, Nangang District, Heilongjiang, Harbin Patentee before: Heilongjiang University |