CN110982860A - Fermentation method of curdlan - Google Patents
Fermentation method of curdlan Download PDFInfo
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- CN110982860A CN110982860A CN201911256701.0A CN201911256701A CN110982860A CN 110982860 A CN110982860 A CN 110982860A CN 201911256701 A CN201911256701 A CN 201911256701A CN 110982860 A CN110982860 A CN 110982860A
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- C12P19/00—Preparation of compounds containing saccharide radicals
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Abstract
The invention relates to a microbial fermentation process, in particular to a fermentation method of curdlan, which comprises the following steps: the method comprises the following steps: inoculating the slant strain preparation bacterial suspension into a seed tank liquid culture medium, and culturing until the slant strain preparation bacterial suspension is mature; step two: culturing mature seeds and transferring the seeds into a culture medium of a fermentation tank to enable thalli to synthesize curdlan outside cells; the scheme improves the conversion rate of the substrate sugar to 70-80%, which is higher than the average level in China by nearly 20%, thereby greatly reducing the production cost and having high gel strength of the finished product.
Description
Technical Field
The invention relates to a microbial fermentation process, in particular to a fermentation method of curdlan.
Background
The curdlan, also known as thermogel, is a biological polysaccharide synthesized extracellularly during growth and reproduction by culturing M.tumefaciens in a fermentation medium.
There are many specific properties of curdlan, which forms thermally irreversible gels, and has edible and various industrial uses. In 1989, japanese korea began to use it as a food gum. The FDA in the united states approved in 1996 for use as a stabilizer, thickener, in food ingredients. The coagulated polysaccharide becomes a food polysaccharide produced by fermentation approved by the FDA after xanthan gum and gellan gum, which provides a wider space for further popularization and application of the coagulated polysaccharide. The application of the curdlan and the development of food also reach a new level. It has been produced in japan, canada, etc., and has been developed and applied to many foods in japan and taiwan of china.
At present, the production of curdlan in China has been on an initial scale, products of companies such as Shandong Chinese family organisms, Hebei Xinhe organisms and the like are accepted by markets, but the problems of low conversion rate of substrate sugar, small capacity, low comprehensive equipment utilization rate and the like still exist in the production process.
Disclosure of Invention
In order to solve the above problems in the prior art, the present invention provides a fermentation method of curdlan. The technical problem to be solved by the invention is realized by the following technical scheme: the method comprises the following steps:
the method comprises the following steps: inoculating the slant strain preparation bacterial suspension into a seed tank liquid culture medium, and culturing until the slant strain preparation bacterial suspension is mature;
step two: culturing mature seeds and transferring the seeds into a culture medium of a fermentation tank, so that the thalli synthesize curdlan outside cells.
Further, the formula of the liquid culture medium is 20g/L of glucose, 10g/L of yeast powder, 20g/L of peptone and 0.15g/L of antifoaming agent.
Further, the formula of the fermentation tank culture medium in the second step is as follows: 30g/L of carbon source, 5g/L of yeast powder, 2g/L of diammonium hydrogen phosphate, 1g/L of monopotassium phosphate, 0.2g/L of magnesium sulfate and 0.1g/L of defoaming agent.
Further, the carbon source is a mixture of sucrose and starch hydrolysate.
Further, the preparation method of the starch hydrolysate comprises the steps of firstly adding water with the volume calculated by 1/2 of fermentation medium, adding corn starch, heating to 80-90 ℃, gelatinizing for 30min, and adding α -amylase according to 0.05 per mill of the mass of the starch for liquefying for 1 min.
Further, the mature culture in the first step comprises the steps of conducting air digestion of a ① fermentation tank at 0.15-0.18 MPa, maintaining pressure for 30 minutes, conducting actual digestion of a ② fermentation tank, namely putting a liquid seed culture medium into the tank, metering the liquid seed culture medium according to 70% of the volume of the fermentation tank, adjusting the pH value to 6.0, closing a tank cover, heating a jacket to 90 ℃, directly heating the liquid seed culture medium to 121 ℃ by steam, keeping the temperature at 0.12MPa for 20 minutes, cooling ③, cooling to 28 +/-2 ℃ after sterilization, allowing inoculation, inoculating bacterial suspension in ④ by a zero pound method, and conducting ⑤ culture at 28 +/-2 ℃, 0.05MPa of tank pressure, 0.2-0.8 VVM of air volume and stirring speed150-250 rpm, dissolved oxygen of more than 30%, culture time of 15-24 hours, OD600Can be rotated 10-15 times.
Further, the step of transferring the strain into a fermentation tank comprises the following steps of ① empty tank sterilization, wherein the pressure is 0.15-0.18 MPa, the pressure is maintained for 30 minutes, ② solid tank sterilization, fermentation medium is put into the tank, the material is counted according to 50% of the volume of the fermentation tank, the tank cover is closed, a jacket is preheated to 85-95 ℃, steam is heated to 121 ℃, the pressure is 0.12MPa, the temperature is kept for 20 minutes, the temperature is cooled to 28 +/-2 ℃, the tank pressure is 0.05MPa, the pH is adjusted to 6.8 +/-0.2 for inoculation, ③ inoculation is to inoculate the seed solution into the tank for cultivation by adopting a pressure difference method, the temperature is ④ cultivation, the temperature is 28 +/-2 ℃, the tank pressure is 0.05MPa, the air volume is 0.6-1.2 VVM, the pH is 5.5, the stirring speed is 80-150 rpm, and the.
Compared with the prior art, the invention has the beneficial effects that:
according to the invention, the carbon source in the fermentation medium is adjusted, so that the conversion rate of the substrate sugar is improved to 70-80% which is higher than the average level in China by nearly 20%, thus the production cost is greatly reduced, and the gel strength of the finished product is higher than the national standard GB 28304-2012.
Detailed Description
The present invention will be described in further detail with reference to specific examples, but the embodiments of the present invention are not limited thereto.
Example 1
A fermentation method of curdlan comprises culturing strain in liquid culture medium to obtain mature seed, transferring into the fermentation culture medium, and fermenting to obtain curdlan;
wherein the formula (g/L) of the liquid culture medium is as follows: glucose 20, yeast powder 10, peptone 20 and an antifoaming agent 0.15;
wherein the formula (g/L) of the fermentation medium is as follows: carbon source 30 (wherein sucrose 5, starch hydrolysate 25), yeast powder 5, diammonium hydrogen phosphate 2, potassium dihydrogen phosphate 1, magnesium sulfate 0.2, and defoaming agent 0.1;
the preparation method of the starch hydrolysate in the fermentation medium comprises the steps of firstly adding water with the volume calculated by the fermentation medium 1/2 and adding corn starch, heating to 80-90 ℃ for gelatinization for 30min, adding α -amylase according to 0.05 per mill of the mass of the starch for liquefaction for 1min, and then adding other materials.
The culture process of mature seeds comprises the following steps of empty digestion of a ① fermentation tank at 0.15MPa, pressure maintaining for 30 minutes, actual digestion of a ② fermentation tank at 1.4T liquid seed culture medium, putting the fermentation tank at 2T, adjusting pH to 6.0, closing a tank cover, heating a jacket to 92 ℃, directly heating the jacket to 121 ℃ by steam, carrying out heat preservation at 0.12MPa for 20 minutes, cooling ③, cooling to 28 +/-2 ℃ after sterilization, waiting for inoculation, inoculating ④ by a zero pound method, inoculating a fire ring which is dipped in alcohol, sleeving an inoculation cap, slightly unscrewing the inoculation cap, closing an air inlet tank valve, rapidly closing a large exhaust gas, reducing the tank pressure to below 0.01MPa, igniting the fire ring, slightly unscrewing the inoculation cap, placing the fire ring above a flame, slightly pulling a cotton plug beside the flame ring, burning the bottle opening on the flame for 3-5 seconds, rapidly pouring the liquid into the bottle opening, rapidly pouring the bottle opening into the bottle without leaving the flame, rapidly burning the bottle opening on the flame, tightly, removing the fire ring, carrying out the wet culture at a temperature of 150.05-2 rpm, stirring at 0.05-2 ℃ and a rotation speed of 20.05-2M, and stirring for culture at a rotation speed of more than 0.05-2 hours60011.8 rotary pots.
The fermentation culture process includes ① empty tank sterilization under 0.15MPa for 30min, ② solid tank sterilization, including 10T fermentation medium in 20T fermentation tank, closing the tank cover, preheating the tank cover to 90 deg.c, direct steam heating to 121 deg.c and 0.12MPa, maintaining for 20 min, cooling to 28 +/-2 deg.c, 0.05MPa and 6.82 pH, inoculating ③ seed liquid, culturing ④ at 28 +/-2 deg.c, 0.05MPa pressure, 0.6-1.2 VVM and 5.5 pH, stirring speed of 80-150 rpm and dissolved oxygen content of over 30%, ⑤ fermentation liquid with residual sugar concentration of 0.24g/L, substrate sugar conversion rate of 76% and product gel strength of 610g/cm2；
Example 2
A fermentation method of curdlan comprises culturing strain in liquid culture medium to obtain mature seed, transferring into the fermentation culture medium, and fermenting to obtain curdlan;
wherein the formula (g/L) of the liquid culture medium is as follows: glucose 20, yeast powder 10, peptone 20 and an antifoaming agent 0.15;
wherein the formula (g/L) of the fermentation medium is as follows: carbon source 30 (wherein sucrose 7 and starch hydrolysate 23), yeast powder 5, diammonium hydrogen phosphate 2, potassium dihydrogen phosphate 1, magnesium sulfate 0.2 and defoaming agent 0.1;
the preparation method of the starch hydrolysate in the fermentation medium comprises the steps of firstly adding water with the volume calculated by the fermentation medium 1/2 and adding corn starch, heating to 80-90 ℃ for gelatinization for 30min, adding α -amylase according to 0.05 per mill of the mass of the starch for liquefaction for 1min, and then adding other materials.
The culture process of mature seeds comprises the following steps of empty digestion of a ① fermentation tank at 0.15MPa, pressure maintaining for 30 minutes, actual digestion of a ② fermentation tank at 1.4T liquid seed culture medium, putting the fermentation tank at 2T, adjusting pH to 6.0, closing a tank cover, heating a jacket to 92 ℃, directly heating the jacket to 121 ℃ by steam, carrying out heat preservation at 0.12MPa for 20 minutes, cooling ③, cooling to 28 +/-2 ℃ after sterilization, waiting for inoculation, inoculating ④ by a zero pound method, inoculating a fire ring which is dipped in alcohol, sleeving an inoculation cap, slightly unscrewing the inoculation cap, closing an air inlet tank valve, rapidly closing a large exhaust gas, reducing the tank pressure to below 0.01MPa, igniting the fire ring, slightly unscrewing the inoculation cap, placing the fire ring above a flame, slightly pulling a cotton plug beside the flame ring, burning the bottle opening on the flame for 3-5 seconds, rapidly pouring the liquid into the bottle opening without leaving the flame, rapidly burning the bottle opening on the flame, tightly, removing the fire ring, carrying out the wet culture at a temperature of 150.05-2 rpm, stirring at 0.05-2.05-2 ℃ and a rotation speed of 0.05-2 rpm, and stirring for culture time of more than 0.05-2M60012.7 rotary pots.
The fermentation culture process includes ① empty sterilization under 0.15MPa for 30min, ② solid sterilization under 20T fermentation medium, closing the fermentation tank, preheating the tank cover to 90 deg.c, direct steam heating to 121 deg.c and 0.12MPa, maintaining for 20 min, cooling to 28 +/-2 deg.c, regulating pH to 6.82 for inoculation, ③ inoculation with pressure difference method to culture the seed liquid in the tank, ④ culture at 28 +/-2 deg.c and 0.05MPa in the tank, and wind blowing to culture the seed liquid in the tank under ④ conditions of 28 +/-2 deg.c and 0.05MPaThe amount of the product is 0.6-1.2 VVM, the ph is 5.5, the stirring speed is 80-150 rpm, the dissolved oxygen is more than 30%, ⑤ when the fermentation is finished, the concentration of residual sugar in the fermentation liquor is 0.28g/L, the conversion rate of substrate sugar is 74%, and the gel strength of the product is 590g/cm2；
Example 3
A fermentation method of curdlan comprises culturing strain in liquid culture medium to obtain mature seed, transferring into the fermentation culture medium, and fermenting to obtain curdlan;
wherein the formula (g/L) of the liquid culture medium is as follows: glucose 20, yeast powder 10, peptone 20 and an antifoaming agent 0.15;
wherein the formula (g/L) of the fermentation medium is as follows: carbon source 30 (wherein sucrose 10 and starch hydrolysate 20), yeast powder 5, diammonium hydrogen phosphate 2, potassium dihydrogen phosphate 1, magnesium sulfate 0.2 and defoaming agent 0.1;
the preparation method of the starch hydrolysate in the fermentation medium comprises the steps of firstly adding water with the volume calculated by the fermentation medium 1/2 and adding corn starch, heating to 80-90 ℃ for gelatinization for 30min, adding α -amylase according to 0.05 per mill of the mass of the starch for liquefaction for 1min, and then adding other materials.
The culture process of mature seeds comprises the following steps of empty digestion of a ① fermentation tank at 0.15MPa, pressure maintaining for 30 minutes, actual digestion of a ② fermentation tank at 1.4T liquid seed culture medium, putting the fermentation tank at 2T, adjusting pH to 6.0, closing a tank cover, heating a jacket to 92 ℃, directly heating the jacket to 121 ℃ by steam, carrying out heat preservation for 20 minutes at 0.12MPa, cooling ③, cooling to 28 +/-2 ℃ after sterilization, waiting for inoculation, inoculating ④ by a zero pound method, inoculating a fire ring which is dipped in alcohol, sleeving an inoculation cap, slightly unscrewing the inoculation cap, closing an air inlet tank valve, rapidly closing a large exhaust gas, reducing the tank pressure to below 0.01MPa, igniting the fire ring, slightly unscrewing the inoculation cap, placing the fire ring above a flame, slightly pulling a cotton plug beside the flame ring, burning the bottle opening on the flame for 3-5 seconds, rapidly pouring liquid into the bottle opening, rapidly pouring the liquid into the bottle opening without leaving the flame, rapidly burning the bottle opening on the flame, rapidly, tightly screwing the fire ring, removing the fire ring, carrying out the wet culture at a temperature of 0.05-2 rpm, stirring at 0.05-2%, carrying out the wet culture time of 0.05-2 rpm, and culturing at a rotation speed of 20.05-2MHour, OD60013.3 rotary pot.
The fermentation culture process includes ① empty tank sterilization under 0.15MPa for 30min, ② solid tank sterilization, including 10T fermentation medium inside 20T fermentation tank, closing the tank cover, preheating the tank cover to 90 deg.c, direct steam heating to 121 deg.c and 0.12MPa, maintaining for 20 min, cooling to 28 +/-2 deg.c, 0.05MPa, regulating pH to 6.82 for inoculation, ③ inoculating seed liquid into tank for culture, ④ culture at 28 +/-2 deg.c, 0.05MPa, 0.6-1.2 VVM and pH 5.5, stirring speed of 80-150 rpm and dissolved oxygen content over 30%, ⑤ fermentation liquid with residual sugar concentration of 0.22g/L, substrate sugar conversion rate of 79% and product gel strength of 640g/cm2；
The foregoing is a more detailed description of the invention in connection with specific preferred embodiments and it is not intended that the invention be limited to these specific details. For those skilled in the art to which the invention pertains, several simple deductions or substitutions can be made without departing from the spirit of the invention, and all shall be considered as belonging to the protection scope of the invention.
Claims (7)
1. A fermentation process for curdlan, characterized by: the method comprises the following steps:
the method comprises the following steps: inoculating the slant strain preparation bacterial suspension into a seed tank liquid culture medium, and culturing until the slant strain preparation bacterial suspension is mature;
step two: culturing mature seeds and transferring the seeds into a culture medium of a fermentation tank, so that the thalli synthesize curdlan outside cells.
2. A fermentation process of curdled polysaccharide according to claim 1, wherein: the formula of the liquid culture medium is 20g/L of glucose, 10g/L of yeast powder, 20g/L of peptone and 0.15g/L of antifoaming agent.
3. A fermentation process of curdled polysaccharide according to claim 1, wherein: the formula of the fermentation tank culture medium in the step two is as follows: 30g/L of carbon source, 5g/L of yeast powder, 2g/L of diammonium hydrogen phosphate, 1g/L of monopotassium phosphate, 0.2g/L of magnesium sulfate and 0.1g/L of defoaming agent.
4. A fermentation process of curdled polysaccharide according to claim 3, wherein: the carbon source is a mixture of sucrose and starch hydrolysate.
5. The fermentation method of coagulated polysaccharide according to claim 4, wherein the preparation method of the starch hydrolysate comprises adding water in a volume calculated by fermentation medium 1/2, adding corn starch, heating to 80-90 deg.C, gelatinizing for 30min, and adding α -amylase at a weight of 0.05 ‰ of starch for liquefaction for 1 min.
6. The fermentation method of curdlan according to claim 1, wherein the fermentation step of culturing and maturing in the first step comprises the steps of conducting empty digestion in a ① fermentation tank at 0.15-0.18 MPa for 30 minutes, conducting actual digestion in a ② fermentation tank, feeding a liquid seed culture medium into the tank, metering 70% of the volume of the fermentation tank, adjusting pH to 6.0, closing a tank cover, heating a jacket to 90 ℃, directly heating the liquid seed culture medium to 121 ℃ by steam, conducting heat preservation at 0.12MPa for 20 minutes, conducting ③ cooling, cooling after sterilization, cooling to 28 +/-2 ℃ for inoculation, conducting inoculation on a bacterial suspension in a zero pound method in ④, and conducting ⑤ culture at 28 +/-2 ℃, 0.05MPa of tank pressure, 0.2-0.8 VVM of air volume, 150-250 rpm of stirring speed, more than 30% of dissolved oxygen, 15-24 hours of culture time and OD600Can be rotated 10-15 times.
7. The fermentation method of curdled polysaccharide according to claim 1, wherein the step of transferring to a fermentation tank comprises ① sterilization in an empty tank under 0.15-0.18 MPa for 30 minutes, ② sterilization in a solid tank, wherein fermentation medium is charged into the tank, 50% of the fermentation tank volume is measured, the tank cover is closed, the jacket is preheated by 85-95 ℃, steam is heated to 121 ℃, the pressure is 0.12MPa, the temperature is maintained for 20 minutes, the temperature is cooled to 28 +/-2 ℃, the tank pressure is 0.05MPa, the pH is adjusted to 6.8 +/-0.2 for inoculation, ③ inoculation is performed by inoculating seed liquid into the tank by a pressure difference method, ④ cultivation is performed at 28 +/-2 ℃, the tank pressure is 0.05MPa, the air flow is 0.6-1.2 VVM, the pH is 5.5, the stirring speed is 80-150 rpm, and the dissolved oxygen is more than 30%.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111621527A (en) * | 2020-05-12 | 2020-09-04 | 浙江兰博生物科技股份有限公司 | Biological fermentation process and device thereof |
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Patent Citations (4)
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| CN102277319A (en) * | 2011-08-02 | 2011-12-14 | 河北鑫合生物化工有限公司 | Rhizobium radiobacter and method for producing curdian fermentation liquor by using strain |
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| CN111621527A (en) * | 2020-05-12 | 2020-09-04 | 浙江兰博生物科技股份有限公司 | Biological fermentation process and device thereof |
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Application publication date: 20200410 |