CN112391421A - Culture method for improving oil content of saccharomyces cerevisiae by adjusting total salt content of saccharomyces cerevisiae fermentation liquor and high-oil-yield saccharomyces cerevisiae - Google Patents
Culture method for improving oil content of saccharomyces cerevisiae by adjusting total salt content of saccharomyces cerevisiae fermentation liquor and high-oil-yield saccharomyces cerevisiae Download PDFInfo
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Abstract
The invention relates to a culture method for improving the oil content of saccharomyces cerevisiae by adjusting the total salt content of saccharomyces cerevisiae fermentation liquor and saccharomyces cerevisiae with high oil yield, belonging to the technical field of biology. The method effectively avoids the defects of huge workload and random and uncertain results of large-scale screening of the oil-producing saccharomyces cerevisiae strains, and simultaneously, compared with the conventional microbial oil accumulation induction method, the method for culturing the saccharomyces cerevisiae can obtain higher oil content and oil yield, can solve the problem of insufficient high-value palmitoleic acid resource amount, has great industrial production and application values, and can bring excellent economic benefit and social benefit.
Description
Technical Field
The invention belongs to the technical field of biology, relates to a culture method for improving the oil content of saccharomyces cerevisiae, and particularly relates to a method for improving the oil content of saccharomyces cerevisiae by adjusting the total salt content in fermentation liquor in the fermentation culture process of saccharomyces cerevisiae.
Background
The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.
Saccharomyces cerevisiae is widely used in brewing industry and pastry food processing industry, and meanwhile, the grease of the Saccharomyces cerevisiae is rich in omega-7 monounsaturated fatty acid, palmitoleic acid (hexadecenoic acid, C16:1), which is very rare and expensive in nature. Palmitoleic acid is proved to have remarkable prevention and treatment effects on diseases such as type II diabetes, metabolic function syndrome, hyperlipidemia, hypertension and the like, and has been developed into a medicinal preparation to be marketed in countries such as Europe and America, but raw materials of the products mainly come from wild animal and plant resources which are difficult to be commercially cultured or planted, such as deep sea fish, sea buckthorn, macadamia nuts and the like, so that the market development is limited. The proportion of palmitoleic acid in total lipid of saccharomyces cerevisiae can reach about 50%, so that the palmitoleic acid has important value and good application prospect in the field of palmitoleic acid production. However, since saccharomyces cerevisiae cannot accumulate a large amount of oil and fat, and usually, the oil and fat content thereof is only about 5% of the dry cell weight, and the oil and fat content thereof rarely exceeds 20% even when it is cultured in an oil and fat induction medium such as nitrogen deficiency, saccharomyces cerevisiae is not generally considered to be an oleaginous microorganism, and there is little research on the oil production of saccharomyces cerevisiae. This is because the organic carbon source is preferentially converted into cell biomass and ethanol by the saccharomyces cerevisiae during the fermentation culture process rather than accumulating oil, so if the saccharomyces cerevisiae is used for oil production, the yield of sugar to oil is very low, and the sugar consumption per unit of oil production is necessarily very high. Meanwhile, the low oil content of the saccharomyces cerevisiae is not beneficial to oil extraction, and the extraction rate is low. These all greatly increase the cost of producing oil by saccharomyces cerevisiae. Therefore, in order to realize the industrial production of the rare palmitoleic acid by using the saccharomyces cerevisiae, a new technical method must be developed to improve the oil content of the yeast.
At present, the saccharomyces cerevisiae is mainly used in brewing industry and wheaten food processing industry, and related research documents for oil accumulation of the saccharomyces cerevisiae are few. The method for improving the oil content of the saccharomyces cerevisiae by optimizing a culture medium, particularly controlling the carbon-nitrogen ratio and the like is the most commonly adopted method. If researchers optimize the culture medium of the saccharomyces cerevisiae by a response surface method experiment, the results show that 15 percent of glucose (w/v, the same below), 0.2 percent of peptone, 0.4 percent of yeast extract powder, 0.471 percent of citric acid and MgSO 4 are found4·7H2O 0.1％，ZnSO4·7H2O 0.2％，CaCl2 0.025％，FeSO4·7H2In an optimized culture medium with 0.005% of O, the grease content of the saccharomyces cerevisiae after 96 hours of culture can reach 14.55% to the maximum. The content of the grease is very low compared with other oil-producing microorganisms, and the grease has no production and application value at all. Research personnel researches the induction culture of a high carbon nitrogen ratio culture medium of saccharomyces cerevisiae, and finds that: the oil content of the saccharomyces cerevisiae is improved to 20 percent by fermenting and culturing the saccharomyces cerevisiae in an optimized culture medium with high organic carbon concentration (50g/L) and low nitrogen concentration (5 mM). This indicates that the enhancement effect of the high carbon-nitrogen ratio oil-induced culture on the oil accumulation of saccharomyces cerevisiae is very limited, and the requirement of industrial oil production cannot be met (for example, the oil content of general oil-producing crops and oil-producing microorganisms is about 40%, and if the oil content is lower than this value, the oil-producing cost performance is not high, and the oil-producing method is not suitable for being used as an oil production way).
In recent years, researchers have attempted to increase the lipid content of saccharomyces cerevisiae by means of genetic engineering. For example, in a research aiming at oil production of saccharomyces cerevisiae, the saccharomyces cerevisiae is subjected to genetic modification to strengthen the oil synthesis way, but the oil content of the saccharomyces cerevisiae is only 11.6 percent after the genetic modification, and the oil yield is extremely low and is only 0.023 g/L. More importantly, the safety of the genetically engineered microorganisms used for producing raw materials such as food is still a big controversial, and the genetically engineered microorganisms are difficult to be commercially applied in a short time.
Disclosure of Invention
Aiming at the technical defects and the technical vacancy, the invention provides a novel method for effectively enhancing the oil accumulation of saccharomyces cerevisiae, namely, the total salt content of the fermentation liquor is maintained at 15-30 g/L by adjusting the total salt content of the fermentation liquor in the culture process of the saccharomyces cerevisiae. Under the condition of the total salt content, the cell growth of the saccharomyces cerevisiae cannot be obviously inhibited, but the activity of enzyme related to ethanol synthesis in the cell can be obviously inhibited, so that the ethanol synthesis of the saccharomyces cerevisiae is inhibited, the activity of enzyme related to fatty acid and oil synthesis in the saccharomyces cerevisiae cell is improved, the ratio of organic carbon source flowing to fatty acid synthesis and oil synthesis is increased, and the oil content in the saccharomyces cerevisiae cell is finally and greatly improved. The method is obviously different from the common method for promoting the accumulation of the microbial oil through the high carbon/nitrogen ratio, which is commonly used in the research field, and has more obvious effect on promoting the accumulation of the saccharomyces cerevisiae oil. Meanwhile, the method has no research report for promoting the oil accumulation of the saccharomyces cerevisiae. Therefore, the method has obvious creativity and novelty and also has great industrial production and application values.
In order to achieve the technical purpose, the invention adopts the following technical scheme:
the invention provides a culture method for improving the grease content of saccharomyces cerevisiae by adjusting the total salt content of saccharomyces cerevisiae fermentation liquor, which comprises the following steps: in the culture process of the saccharomyces cerevisiae, the total salt content of the fermentation culture solution is adjusted to be kept between 15 g/L and 30 g/L.
According to the invention, the carbon metabolism pathway of the saccharomyces cerevisiae is analyzed, and the main reason that the oil content is low is that the saccharomyces cerevisiae can preferentially convert an organic carbon source into ethanol instead of basic constituent unit fatty acid of the oil. Therefore, in order to improve the oil content of the saccharomyces cerevisiae, starting with the distribution of organic carbon sources to the synthesis of ethanol and fatty acid, the organic carbon sources flow to the synthesis of ethanol less and fatty acid more by a regulating means, so that a relatively effective method for enhancing the yeast oil production is formed, and the technical report on the aspect is not available at present.
In a second aspect of the invention, there is provided a high oil-yielding Saccharomyces cerevisiae strain cultured by any one of the above methods.
The oil content of the saccharomyces cerevisiae in the invention reaches about 45% of the dry cell weight, but the common method can only enable the oil content of the saccharomyces cerevisiae to reach less than 20% of the dry cell weight, so the method is more than 2.2 times of the effect which can be achieved by the common method. Meanwhile, the method provided by the invention can enable the cell density of the saccharomyces cerevisiae to reach more than 40g/L, and the general method can only enable the cell density of the saccharomyces cerevisiae to reach about 4g/L (provided by the embodiment of the invention) due to the limiting effect on the growth, so that the oil yield which can be achieved by the method is more than 18g/L through simple calculation, while the oil yield which can be achieved by the general method is less than 0.8g/L, and the oil yield of the method provided by the invention is more than 22 times that of the general method.
In a third aspect of the invention, the application of the high-oil-yield saccharomyces cerevisiae in oil production is provided.
Because the oil content of the saccharomyces cerevisiae can be greatly improved, a simpler oil extraction process can be adopted in the oil extraction process, a higher oil extraction rate can be obtained, the oil extraction cost can be reduced, the conversion rate of an organic carbon source to a target product is improved, and the culture cost is reduced, so that the saccharomyces cerevisiae oil extraction method is expected to be widely applied to oil production.
The invention has the beneficial effects that:
(1) the invention provides a culture method for improving the oil content of saccharomyces cerevisiae, which comprises the following steps: in the culture process of the saccharomyces cerevisiae, the total salt content is maintained at 15 to 30g/L by adjusting the total salt content of the fermentation liquor, and under the condition, the oil content of the saccharomyces cerevisiae can reach about 45 percent of the dry weight of cells.
(2) The method provided by the invention has very obvious difference with the common method for promoting the accumulation of the microbial oil through the high carbon-nitrogen ratio in the research field, and the action principle is completely different. The general method has the action principle that the lack of nitrogen element in the culture solution inhibits the growth of microorganisms and the synthesis of protein, thereby reducing the proportion of the protein in the dry cell weight, and therefore, the contents of other two main components, namely sugar and grease, in the dry cell weight are increased relatively. However, the nitrogen deficiency conditions in the conventional methods greatly reduce the cell density (i.e., biomass) of microorganisms in the culture broth, resulting in a decrease in oil production and oil productivity. The method provided by the invention has the action principle that the total salt content of the fermentation liquor is maintained at 15-30 g/L by adjusting the total salt content of the fermentation liquor, and under the condition of the total salt content: on one hand, the cell growth of the saccharomyces cerevisiae is not seriously inhibited, but the activity of enzyme related to ethanol synthesis in the cell is obviously inhibited, so that the ethanol amount produced by saccharomyces cerevisiae fermentation is reduced, namely the proportion of the organic carbon source flowing to ethanol synthesis in the saccharomyces cerevisiae cell is reduced; on the other hand, under the condition of the total salt content, the activity of enzymes related to fatty acid and oil synthesis in the saccharomyces cerevisiae cell is improved, so that the ratio of organic carbon source flow to fatty acid synthesis and oil synthesis is increased, and finally, the oil content in the saccharomyces cerevisiae cell is greatly improved. In the conventional culture of the saccharomyces cerevisiae, the total salt in the culture solution usually only contains nutrient salts such as phosphate and sulfate which are necessary for the growth of the saccharomyces cerevisiae, and the total salt content is very low and is only about 2g/L, but the oil content of the saccharomyces cerevisiae cultured under the condition of the total salt content is very low and is only about 5 percent of the dry weight of cells. Therefore, the method provided by the invention is completely different from the common method, and the common method has no inspiration or reference effect on the method provided by the invention. Therefore, the method provided by the invention has extremely obvious creativity.
(3) From the effect of promoting the oil accumulation, the method provided by the invention can enable the oil content of the saccharomyces cerevisiae to reach about 45% of the dry cell weight, and the general method can only enable the oil content of the saccharomyces cerevisiae to reach less than 20% of the dry cell weight, so the method has the effect more than 2.2 times of that of the general method. Meanwhile, the method provided by the invention can enable the cell density of the saccharomyces cerevisiae to reach more than 40g/L, and the general method can only enable the cell density of the saccharomyces cerevisiae to reach about 4g/L (provided by the embodiment of the invention) due to the limiting effect on the growth, so that the oil yield which can be achieved by the method is more than 18g/L through simple calculation, while the oil yield which can be achieved by the general method is less than 0.8g/L, and the oil yield of the method provided by the invention is more than 22 times that of the general method. In another study aimed at oil production from Saccharomyces cerevisiae, although Saccharomyces cerevisiae was genetically modified to enhance its oil synthesis pathway, its oil content was only 11.6%, and oil yield was only 0.023g/L (Kyu, et al. development of a Saccharomyces cerevisiae strain for creating the interaction of a microbial oil feed for a biodiesel production, 2012,110 (343-. This, on the one hand, again illustrates the difficulty of oil accumulation in saccharomyces cerevisiae and, on the other hand, highlights the beneficial effects of the invention. Meanwhile, the method provided by the invention has not been reported in grease accumulation research of saccharomyces cerevisiae. Therefore, the method provided by the invention has extremely remarkable novelty.
(4) The method provided by the invention can also avoid the defects of large workload, randomness and uncertainty of results in large-scale screening of the oil-producing saccharomyces cerevisiae strains, and the method has wide applicability to the saccharomyces cerevisiae strains.
(5) According to the invention, the oil content of the saccharomyces cerevisiae can be greatly improved, so that a simpler oil extraction process can be adopted in the oil extraction process, a higher oil extraction rate can be obtained, the oil extraction cost can be reduced, the conversion rate of an organic carbon source to a target product is improved, and the culture cost is reduced.
(6) The grease of the saccharomyces cerevisiae is rich in very rare and expensive palmitoleic acid, and has remarkable prevention and treatment effects on diseases such as diabetes, metabolic function syndrome, hyperlipidemia, hypertension and the like. However, at present, palmitoleic acid is mainly derived from wild animal and plant resources which are difficult to be commercially cultured or planted, such as deep sea fish, sea buckthorn, macadamia nut and the like, and the resource amount is extremely limited. Palmitoleic acid has been developed into pharmaceutical preparations to be marketed in countries such as europe and the united states, but market development is limited due to resource problems. In China, the development of diabetes, metabolic function syndrome, hyperlipidemia, hypertension and other diseases is more and more severe, and the development and popularization of the palmitoleic acid related medicinal preparation are limited due to the shortage of resource. The method provided by the invention can exactly solve the problem of insufficient palmitoleic acid resources, has great industrial application value, and also has very important social and economic benefits.
(7) The culture method is simple, high in oil yield, strong in practicability and easy to popularize.
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The accompanying drawings, which are incorporated in and constitute a part of this specification, are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification, illustrate exemplary embodiments of the invention and together with the description serve to explain the invention and not to limit the invention.
FIG. 1 is a graph showing the effect of different total salt contents on growth and oil content of Saccharomyces cerevisiae in example 1 of the present invention;
FIG. 2 is a comparison of the effect of 30g/L total salt content and nitrogen deficiency treatment on Saccharomyces cerevisiae growth and oil content in example 2 of the present invention;
FIG. 3 is a graph showing the effect of total salt content of 30g/L on growth and oil content of Saccharomyces cerevisiae under anaerobic and aerobic conditions in example 3 of the present invention;
FIG. 4 is a graph showing the effect of adjusting the total salt content to 30g/L on the oil content of Saccharomyces cerevisiae at the beginning of different culture stages in example 4 of the present invention;
FIG. 5 shows the oil content of different Saccharomyces cerevisiae strains in example 5 of the present invention under the condition of total salt content of 30 g/L;
FIG. 6 shows the oil content of Saccharomyces cerevisiae adjusted by different salts under the condition of total salt content of culture solution of 30g/L in example 6 of the present invention;
FIG. 7 shows the fatty acid composition of Saccharomyces cerevisiae at a total salt content of 30g/L in example 7 of the present invention.
Detailed Description
It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
As mentioned above, the oil content of the saccharomyces cerevisiae is low, and the effect of promoting the oil accumulation of the saccharomyces cerevisiae in the prior art is extremely limited, so that the requirement of mass production of the oil is far from being met. Meanwhile, the saccharomyces cerevisiae grease is rich in palmitoleic acid with important medical value, and the mass production of the grease rich in palmitoleic acid has extremely important social and economic benefits. Therefore, a rapid and effective method for promoting the mass accumulation of the saccharomyces cerevisiae grease is urgently needed to be invented.
In view of the above, the present invention provides a method for greatly increasing the oil content of saccharomyces cerevisiae, the method comprising: during the culture process of the saccharomyces cerevisiae, the total salt content is maintained between 15 and 30g/L by adjusting the total salt content in the culture solution, so as to promote the accumulation of saccharomyces cerevisiae grease.
The invention aims to provide a culture method for improving the oil content of saccharomyces cerevisiae.
The second purpose of the invention is to provide the saccharomyces cerevisiae grease prepared by the method.
In a first aspect of the invention, a culture method for increasing the oil content of saccharomyces cerevisiae is provided, and the method comprises the following steps: during the culture process of the saccharomyces cerevisiae, the total salt content is maintained between 15 and 30g/L by adjusting the total salt content in the culture solution, so as to promote the accumulation of saccharomyces cerevisiae grease.
In some embodiments, the Saccharomyces cerevisiae english name is Saccharomyces cerevisiae, also known as baker's yeast, or budding yeast, or colloquially as brewer's yeast, or wine yeast, or white wine yeast, or wine yeast;
in some embodiments, the culture process comprises an anaerobic fermentation culture process and/or an aerobic fermentation culture process;
in some embodiments, the process of adjusting the total salt content in the culture medium to be maintained between 15 and 30g/L can be started at any stage of the whole culture process;
in some embodiments, the salts used to adjust the total salt content of the culture medium include, but are not limited to, one or more of sodium salt, potassium salt, calcium salt, magnesium salt, iron salt, sea salt, and/or a mixture thereof mixed in any ratio.
In a second aspect of the invention, the saccharomyces cerevisiae grease prepared by the method is provided.
The invention is further illustrated by the following examples, which are not to be construed as limiting the invention thereto. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The test methods in the following examples, which are not specified under specific conditions, are generally carried out under conventional conditions. Meanwhile, the saccharomyces cerevisiae used in the embodiment of the invention is a recorded wild species, and can be purchased from commercial channels or separated and purified from natural environment by ordinary technicians, and biological material preservation for patent procedures is not needed.
In the following examples, the Saccharomyces cerevisiae used in examples 1-4, 6 and 7 was designated SC-17 and purchased from Saccharomyces fumaraensis, Inc. under the trade designation S-23.
Example 1
The growth and the oil content of the saccharomyces cerevisiae under the condition of different total salt contents comprise the following specific steps:
the total salt content in the culture solution of the saccharomyces cerevisiae is respectively 1, 2, 5, 10, 15, 20, 30 and 40g/L by adding sodium chloride, and the saccharomyces cerevisiae is cultured by using the culture media with different total salt contents. After continuous culture for 4 days, the biomass of Saccharomyces cerevisiae in the culture medium (gravimetric method) and its oil content (chloroform-methanol method) were determined. Other culture conditions were: the inoculation amount is 5 percent, the glucose in the culture medium is 150g/L, the yeast extract powder is 1 percent, the peptone is 2 percent, the temperature is 28 ℃, the rotating speed is 200rpm, the air ventilation amount is 0.5vvm, and the pH is 6.
As shown in FIG. 1, under the condition that the total salt content is 15-30 g/L, the biomass of the Saccharomyces cerevisiae can reach more than 40g/L, and the oil content is about 45% of the dry weight of the cells. Under the conventional culture condition with lower total salt content (such as 2g/L), the biomass of the saccharomyces cerevisiae can only reach about 35g/L, and the oil content is only about 5 percent of the dry weight of cells, and the calculation shows that the oil yield of the saccharomyces cerevisiae can reach about 10.3 times of that under the conventional culture condition of the saccharomyces cerevisiae by adjusting the total salt content of the culture solution to be between 15 and 30 g/L.
Example 2
The method comprises the following specific steps of comparing the influence of the total salt content condition of 30g/L and the nitrogen deficiency on the growth and the oil content of the saccharomyces cerevisiae:
the total salt content in the culture solution of the saccharomyces cerevisiae is 30g/L by adding sodium chloride, and the saccharomyces cerevisiae is cultured by using the culture medium. After continuous culture for 4 days, the biomass of Saccharomyces cerevisiae in the culture medium (gravimetric method) and its oil content (chloroform-methanol method) were determined. Other culture conditions were: the inoculation amount is 5 percent, the glucose in the culture medium is 150g/L, the yeast extract powder is 3 percent, the peptone is 3 percent, the temperature is 28 ℃, the rotating speed is 200rpm, the air ventilation amount is 0.01vvm, and the pH is 6. Meanwhile, a culture group (without addition of yeast extract and peptone) having a total salt content of 2g/L and a nitrogen deficiency was used as a control.
As shown in FIG. 2, under the cultivation condition of total salt content of 30g/L, the biomass of Saccharomyces cerevisiae can reach 42g/L, and the oil content is 46% of the dry weight of the cells. In a general culture method for inducing microbial oil production, namely a nitrogen deficiency culture group, the biomass of the saccharomyces cerevisiae is only 4g/L, the oil content is 18%, and the oil yield of the saccharomyces cerevisiae under the culture condition of 30g/L total salt content is 26.8 times of the oil yield obtained by a common microbial oil production inducing method.
Example 3
The influence of the total salt content of 30g/L on the growth and the oil content of the saccharomyces cerevisiae under the anaerobic culture condition and the aerobic culture condition comprises the following specific steps:
the total salt content in the culture solution of the saccharomyces cerevisiae is 30g/L by adding sodium chloride, and the saccharomyces cerevisiae is cultured by using the culture medium. After continuous culture for 4 days, the biomass of Saccharomyces cerevisiae in the culture medium (gravimetric method) and its oil content (chloroform-methanol method) were determined. Other culture conditions were: the inoculation amount is 20 percent, the glucose in the culture medium is 150g/L, the yeast extract powder is 6 percent, the peptone is 0 percent, the temperature is 37 ℃, the rotating speed is 5rpm, the pH value is 5, the air ventilation amount is 0.5vvm under aerobic condition, and air is not introduced under anaerobic condition.
As shown in FIG. 3, the total salt content of the culture solution was adjusted to 30g/L under aerobic conditions, the biomass of Saccharomyces cerevisiae could reach 42g/L, and the oil content was 46% of the dry cell weight. Under the anaerobic condition, the total salt content of the culture solution is adjusted to be 30g/L, the biomass of the saccharomyces cerevisiae is adjusted to be 32g/L, and the oil content is adjusted to be 42%. Therefore, the biomass and the grease accumulation of the saccharomyces cerevisiae can still reach a high level under the anaerobic condition. As will be appreciated by those skilled in the art, under anaerobic conditions, the oil accumulation in Saccharomyces cerevisiae is very low, only about 5% of the dry weight of the cells. Therefore, the method can remarkably promote the accumulation of the grease of the saccharomyces cerevisiae under the anaerobic and aerobic culture conditions of the saccharomyces cerevisiae, and has great beneficial effects.
Example 4
The method comprises the following specific steps of adjusting the influence of the total salt content of 30g/L on the oil content of saccharomyces cerevisiae at different culture stages:
in the process of culturing the saccharomyces cerevisiae, the total salt content in a saccharomyces cerevisiae culture solution is made to be 30g/L by adding sodium chloride at different growth stages of the saccharomyces cerevisiae, and the biomass (gravimetric method) and the oil content (chloroform-methanol method) of the saccharomyces cerevisiae in the culture solution are measured after the saccharomyces cerevisiae is continuously cultured for 4 days by using the culture medium. Other culture conditions were: the inoculation amount is 1 percent, the glucose in the culture medium is 350g/L, the yeast extract powder is 0 percent, the peptone is 6 percent, the temperature is 28 ℃, the rotating speed is 200rpm, the air ventilation amount is 0.5vvm, and the pH is 3. Wherein the different culturing stages comprise: lag phase, logarithmic phase, stationary phase, and decline phase.
As shown in FIG. 4, the oil and fat content of the yeast can be increased to about 45% by continuously culturing the Saccharomyces cerevisiae while adjusting the total salt content of the culture solution to 30g/L at different growth stages of the yeast.
Example 5
The oil content of different saccharomyces cerevisiae strains under the condition of total salt content of 30g/L comprises the following specific steps:
culturing 40 strains of Saccharomyces cerevisiae purchased from Angel Yeast (SY wine yeast, RW wine yeast, ordinary yeast, low-sugar yeast, high-sugar-resistant yeast, white wine yeast) under the condition of 30g/L of total salt content in the culture solution, wherein SC 1-SC 6 are purchased from Angel Yeast company (S-33, BE-134, S-04, US-05, BE-256, WB-06, T-58, K-97, W-34/70, S-189, S-23), SC 18-SC 32 are purchased from Rhododeng Yeast company (M44, M05, M21, M76, M639, M20, M02, M29, M42, M47, M41, M54, M84, and SC 84 are purchased from Israel Yeast company (S84-SC 84), and SC 84 are purchased from Israel Yeast, Saisen, Munich, Belgium house, Diamond, West coast BRY-97, London ESD, Nortinham). During the culture, the total salt content in the culture solution was adjusted to 30g/L by adding sodium chloride, and the oil content of Saccharomyces cerevisiae in the culture solution was measured after continuous culture for 4 days (chloroform-methanol method). Other culture conditions were: the inoculation amount is 5 percent, the glucose content in the culture medium is 50g/L, the yeast extract powder is 1 percent, the peptone content is 2 percent, the temperature is 20 ℃, the rotating speed is 600rpm, the air ventilation amount is 0.5vvm, and the pH value is 6.
The results are shown in FIG. 5, 40 Saccharomyces cerevisiae strains with different numbers are cultured under the condition that the total salt content of the culture solution is 30g/L, and the oil content of the Saccharomyces cerevisiae strains can reach about 45%. Therefore, the method has wide applicability to the saccharomyces cerevisiae strains.
Example 6
Adjusting the oil content of the saccharomyces cerevisiae under the condition that the total salt content of the culture solution is 30g/L by using different salts, and specifically comprising the following steps:
a plurality of culture media with total salt content of 30g/L are prepared by respectively adding 30g/L of sodium chloride, 30g/L of potassium chloride, 25.8g/L of sodium chloride, 2g/L of calcium chloride, 2g/L of magnesium chloride, 0.2g/L of ferric chloride (recorded as mixed salt) and 30g/L of sea salt, and the saccharomyces cerevisiae is cultured by the culture media. After continuous culture for 4 days, the oil content of Saccharomyces cerevisiae in the culture solution was measured (chloroform-methanol method). Other culture conditions were: the inoculation amount is 5 percent, the glucose in the culture medium is 150g/L, the yeast extract powder is 2 percent, the peptone is 4 percent, the temperature is 12 ℃, the rotating speed is 1000rpm, the air ventilation amount is 3vvm, and the pH value is 6.8.
As shown in FIG. 6, the oil content of the cultured Saccharomyces cerevisiae was about 45% when the total salt content of the culture solution was adjusted to 30g/L with different salts.
Example 7
The fatty acid composition of the saccharomyces cerevisiae under the condition of total salt content of 30g/L comprises the following specific steps:
the total salt content in the culture solution of the saccharomyces cerevisiae is 30g/L by adding sodium chloride, and the saccharomyces cerevisiae is cultured by using the culture medium. After continuous culture for 4 days, the fatty acid composition of the oil and fat of Saccharomyces cerevisiae in the culture broth was measured (gas chromatography). Other culture conditions were: the inoculation amount is 5 percent, the glucose in the culture medium is 150g/L, the yeast extract powder is 1 percent, the peptone is 2 percent, the temperature is 28 ℃, the rotating speed is 200rpm, the air ventilation amount is 0.5vvm, and the pH is 6.
As shown in FIG. 7, the fatty acid composition of the Saccharomyces cerevisiae oil was mainly palmitoleic acid (C16:1), and it accounted for about 50% of the total fatty acids. As described above, palmitoleic acid is a very rare and expensive monounsaturated fatty acid, which has very important application value in the medical field. The method provided by the invention can greatly improve the grease content of the saccharomyces cerevisiae rich in palmitoleic acid, can solve the problem of insufficient palmitoleic acid resources, has great industrial application value, and also has very important social and economic benefits.
It should be noted that the above-mentioned embodiments are only preferred embodiments of the present invention, and the present invention is not limited thereto, and although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications and equivalents can be made in the technical solutions described in the foregoing embodiments, or equivalents thereof. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention. Although the present invention has been described with reference to the specific embodiments, it should be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention.
Claims (10)
1. A culture method for improving the oil content of saccharomyces cerevisiae by adjusting the total salt content of saccharomyces cerevisiae fermentation liquor is characterized by comprising the following steps: in the culture process of the saccharomyces cerevisiae, the total salt content of the fermentation culture solution is adjusted to be kept between 15 g/L and 30 g/L.
2. The method of claim 1, wherein the Saccharomyces cerevisiae is Saccharomyces cerevisiae and is a strain or a mixture of strains.
3. The method for increasing the oil content of saccharomyces cerevisiae by adjusting the total salt content of the saccharomyces cerevisiae fermentation broth according to claim 1, wherein the culturing process comprises: an anaerobic fermentation culture process and/or an aerobic fermentation culture process.
4. The method of claim 1, wherein the salts in the culture medium comprise: sodium salt, potassium salt, calcium salt, magnesium salt, iron salt, and sea salt.
5. The culture method for increasing the oil content of saccharomyces cerevisiae by adjusting the total salt content of saccharomyces cerevisiae fermentation broth according to claim 1, wherein the inoculation amount is 1-20% in the culture process.
6. The culture method for increasing the oil content of saccharomyces cerevisiae by adjusting the total salt content of saccharomyces cerevisiae fermentation broth according to claim 1, wherein the culture medium further comprises 50-350 g/L of glucose, 0-6% of yeast extract powder and 0-6% of peptone.
7. The culture method for increasing the oil content of saccharomyces cerevisiae by adjusting the total salt content of saccharomyces cerevisiae fermentation liquor according to claim 6, wherein the culture conditions are that the temperature is 12-37 ℃, the rotation speed is 5-1000 rpm, the air ventilation is 0.01-3 vvm or air is not introduced, and the pH is 3-6.8.
8. The method of claim 1, further comprising collecting the saccharomyces cerevisiae cells at the end of the culturing to extract the oil.
9. An oil-producing s.cerevisiae yeast cultured by the method of any one of claims 1-8.
10. Use of the high oil producing saccharomyces cerevisiae according to claim 9 in oil production.
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| CN116478841A (en) * | 2023-05-31 | 2023-07-25 | 中国科学院青岛生物能源与过程研究所 | Saccharomyces cerevisiae mutant strain MU310 capable of producing grease at high yield and application thereof |
| CN115521951B (en) * | 2022-10-12 | 2024-05-31 | 中国科学院青岛生物能源与过程研究所 | Method for accumulating grease by performing induction culture on waste saccharomyces cerevisiae mud |
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| CN116478841A (en) * | 2023-05-31 | 2023-07-25 | 中国科学院青岛生物能源与过程研究所 | Saccharomyces cerevisiae mutant strain MU310 capable of producing grease at high yield and application thereof |
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