The invention relates to 2apos; fucosyllactose fermentation medium

Abstract

The invention discloses a 2 '-fucosyllactose fermentation culture medium, belongs to the technical field of 2'-fucosyllactose, and aims to solve the problem that a microbial whole-cell synthesis method of 2 '-FL has the characteristics of mild conditions, low cost and the like and is the most common 2'-FL synthesis method, the fermentation culture medium is further optimized, effective synthesis of 2 '-FL with glucose and lactose as substrates is realized, and the yield of 2'-fucosyllactose is increased. In order to solve the problem of providing a new strategy for biological preparation of 2 '-FL, the culture medium comprises 20-30 g / L of corn steep liquor dry powder, 18-25 g / L of yeast extract powder, 10-15 g / L of K2HPO4. 3H2O, 2-5 g / L of KH2PO4, 3-8 g / L of urea, 1-5 g / L of MgSO4 (anhydrous), 2-6 g / L of KIO3, 5-20 g of glucose and 20-45 g of lactose. According to the method, KIO3 is added on the basis of an original fermentation culture medium, yeast peptone in original culture is replaced with corn steep liquor dry powder, the content of 2 '-FL produced by metabolism of strains in the optimized fermentation culture medium is much higher than the metabolic content of 2'-FL in the original fermentation culture medium, and meanwhile, the time for stopping rising of the biomass of the strains is delayed to 40 h.

Description

Technical Field

[0001] The invention belongs to the technical field of 2'-fucosyllactose, and in particular relates to a 2'-fucosyllactose fermentation medium. Background Art

[0002] Human milk oligosaccharides (HMOs) are the third largest component in breast milk after lactose and fat, and are an important component for protecting the health of newborns. 2'-fucosyllactose (2'-FL) has the highest content in HMOs, reaching 31%. It is also the simplest oligosaccharide and has most of the functions of HMOs. Therefore, exploring how to synthesize 2'-FL safely and efficiently is of great significance to promoting the industrialization of 2'-FL.

[0003] The microbial whole-cell synthesis of 2'-FL has the characteristics of mild conditions and low cost, and is the most commonly used method for synthesizing 2'-FL. Further optimization of the fermentation medium can achieve the effective synthesis of 2'-FL with glucose and lactose as substrates, providing a new strategy for the biological preparation of 2'-FL. To this end, we proposed a 2'-fucosyllactose fermentation medium. Summary of the invention

[0004] The object of the present invention is to provide a 2'-fucosyllactose fermentation medium to solve the problems raised in the above background technology.

[0005] To achieve the above object, the present invention provides the following technical solution: a 2'-fucosyllactose fermentation medium, characterized in that the medium comprises 20-30 g / L corn steep liquor dry powder, 18-25 g / L yeast extract powder, 10-15 g / L K2HPO4·3H2O, 2-5 g / L KH2PO4, 3-8 g / L urea, 1-5 g / L MgSO4 (anhydrous), 2-6 g / L KIO3, 5-20 g glucose and 20-45 g lactose.

[0006] It should be noted in the scheme that the culture medium includes 20 g / L corn steep liquor powder, 18 g / L yeast extract powder, 10 g / L K2HPO4·3H2O, 2 g / L KH2PO4, 3 g / L urea, 1 g / L MgSO4 (anhydrous), 2 g / L KIO3, 5 g glucose, and 20 g lactose.

[0007] It is further worth noting that the culture medium includes 22 g / L corn steep liquor powder, 20 g / L yeast extract powder, 11 g / L K2HPO4·3H2O, 2-5 g / L KH2PO4, 4 g / L urea, 2 g / L MgSO4 (anhydrous), 33 g / L KIO3, 8 g glucose and 25 g lactose.

[0008] It should be further explained that the culture medium includes 25 g / L corn steep liquor powder, 21 g / L yeast extract powder, 12 g / L K2HPO4·3H2O, 4 g / L KH2PO4, 5 g / L urea, 3 g / L MgSO4 (anhydrous), 4 g / L KIO3, 12 g glucose and 30 g lactose.

[0009] As a preferred embodiment, the culture medium includes 28 g / L corn steep liquor powder, 23 g / L yeast extract powder, 14 g / L K2HPO4·3H2O, 4 g / L KH2PO4, 7 g / L urea, 4 g / L MgSO4 (anhydrous), 5 g / L KIO3, 16 g glucose and 38 g lactose.

[0010] As a preferred embodiment, the culture medium includes 30 g / L corn steep liquor powder, 25 g / L yeast extract powder, 15 g / L K2HPO4·3H2O, 5 g / L KH2PO4, 8 g / L urea, 5 g / L MgSO4 (anhydrous), 6 g / L KIO3, 20 g glucose and 45 g lactose.

[0011] Compared with the prior art, the 2'-fucosyllactose fermentation medium provided by the present invention has at least the following beneficial effects: (1) In the present invention, KIO3 is added to the original fermentation medium, and the yeast peptone in the original culture is replaced with corn steep liquor powder. The content of 2'-FL produced by the metabolism of the strain in the optimized fermentation medium is much higher than that of 2'-FL in the original fermentation medium, and the time for the strain biomass to stop increasing is delayed to 40 h. DETAILED DESCRIPTION

[0012] It should be noted that the references to "one embodiment", "embodiment", "exemplary embodiments", "some embodiments" and the like in the specification indicate that the embodiments described may include specific features, structures or characteristics, but not every embodiment may include the specific features, structures or characteristics. In addition, when a specific feature, structure or characteristic is described in conjunction with an embodiment, it should be within the knowledge of a person skilled in the art to implement such feature, structure or characteristic in conjunction with other embodiments (whether or not explicitly described).

[0013] In general, a term can be understood, at least in part, from its use in context. For example, depending, at least in part, on the context, the term "one or more" as used herein can be used to describe any feature, structure, or characteristic in the singular sense, or can be used to describe a combination of features, structures, or characteristics in the plural sense. Additionally, the term "based on" can be understood as not necessarily intended to convey an exclusive set of factors, but can instead, depending, at least in part, on the context, allow for the presence of other factors that are not necessarily explicitly described.

[0014] It will be understood that the meanings of “on,” “over,” and “above” in the present invention should be interpreted in the broadest manner, so that “on” not only means “directly on” something, but also includes the meaning of being “on” something with intervening features or layers therebetween, and “on” or “over” not only means “on” or “above” something, but also includes the meaning of being “on” or “above” something with no intervening features or layers therebetween.

[0015] The following examples are used to illustrate the present invention, but they cannot be used to limit the scope of protection of the present invention. The conditions in the examples can be further adjusted according to specific conditions. Simple improvements to the method of the present invention under the premise of the concept of the present invention belong to the scope of protection claimed in the present invention.

[0016] The invention provides a 2'-fucosyllactose fermentation medium, which comprises 20-30 g / L corn steep liquor dry powder, 18-25 g / L yeast extract powder, 10-15 g / L K2HPO4·3H2O, 2-5 g / L KH2PO4, 3-8 g / L urea, 1-5 g / L MgSO4 (anhydrous), 2-6 g / L KIO3, 5-20 g glucose and 20-45 g lactose.

[0017] During the fermentation process, the pH is controlled at around 7.25 using ammonia water, and the dissolved oxygen is controlled at 40%-50%.

[0018] Initial dissolved oxygen settings: initial fermentation volume 25L, culture at 37℃, ventilation volume 40L / min, initial rotation speed 100rpm, corrected maximum dissolved oxygen; After 25 hours of fermentation, if the pH automatically rises to above 7.3 and below 7.7, there is no need to add acidic substances to regulate under normal fermentation conditions; When the biomass of the strain stops growing after 30 h of fermentation, 2 g / L KIO3 can be added to delay the period when the biomass of the strain stops growing. The total fermentation time was 50 h; The contents of 2'-fucosyllactose produced in the three fermentations were 51.78, 45.69 and 48.12 g / L, respectively, with an average of 48.53 g / L. Compared with the original fermentation medium, the content of 2'-fucosyllactose produced by the strain increased by 1.4951 times.

[0019] Unless otherwise defined, the technical or scientific terms used in the present invention shall have the usual meanings understood by persons with ordinary skills in the field to which the present invention belongs. The words "including" or "comprising" and the like used in the present invention mean that the elements or objects appearing before the word include the elements or objects listed after the word and their equivalents, but do not exclude other elements or objects. The words "connect" or "connected" and the like are not limited to physical or mechanical connections, but may also include electrical connections, whether direct or indirect. "Up", "down", "left", "right", etc. are only used to indicate relative position relationships. When the absolute position of the object being described changes, the relative position relationship may also change accordingly.

[0020] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A 2'-fucosyllactose fermentation medium, characterized in that: The culture medium comprises 20-30 g / L corn steep liquor dry powder, 18-25 g / L yeast extract powder, 10-15 g / L K2HPO4·3H2O, 2-5 g / L KH2PO4, 3-8 g / L urea, 1-5 g / L MgSO4 (anhydrous), 2-6 g / L KIO3, 5-20 g glucose and 20-45 g lactose.

2. The 2'-fucosyllactose fermentation medium according to claim 1, characterized in that: The culture medium comprises 20 g / L corn steep liquor dry powder, 18 g / L yeast extract powder, 10 g / L K2HPO4·3H2O, 2 g / L KH2PO4, 3 g / L urea, 1 g / L MgSO4 (anhydrous), 32 g / L KIO, 5 g glucose and 20 g lactose.

3. The 2'-fucosyllactose fermentation medium according to claim 1, characterized in that: The culture medium comprises 22 g / L corn steep liquor dry powder, 20 g / L yeast extract powder, 11 g / L K2HPO4·3H2O, 2-5 g / L KH2PO4, 4 g / L urea, 2 g / L MgSO4 (anhydrous), 3 g / L KIO3, 8 g glucose and 25 g lactose.

4. The 2'-fucosyllactose fermentation medium according to claim 1, characterized in that: The culture medium comprises 25 g / L corn steep liquor dry powder, 21 g / L yeast extract powder, 12 g / L K2HPO4·3H2O, 4 g / L KH2PO4, 5 g / L urea, 3 g / L MgSO4 (anhydrous), 34 g / L KIO, 12 g glucose and 30 g lactose.

5. The 2'-fucosyllactose fermentation medium according to claim 1, characterized in that: The culture medium comprises 28 g / L corn steep liquor dry powder, 23 g / L yeast extract powder, 14 g / L K2HPO4·3H2O, 4 g / L KH2PO4, 7 g / L urea, 4 g / L MgSO4 (anhydrous), 35 g / L KIO, 16 g glucose and 38 g lactose.

6. The 2'-fucosyllactose fermentation medium according to claim 1, characterized in that: The culture medium comprises 30 g / L corn steep liquor dry powder, 25 g / L yeast extract powder, 15 g / L K2HPO4·3H2O, 5 g / L KH2PO4, 8 g / L urea, 5 g / L MgSO4 (anhydrous), 36 g / L KIO, 20 g glucose and 45 g lactose.