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JP-7855099-B1 - Method for producing glycolic acid using immobilization technology

JP7855099B1JP 7855099 B1JP7855099 B1JP 7855099B1JP-7855099-B1

Abstract

[Problem] To provide a method for producing glycolic acid using immobilization technology. [Solution] The method includes providing a modified strain containing a specific gene sequence, preparing a polyvinyl alcohol bacteria pellet containing the modified strain, and supplying a carbon source to the polyvinyl alcohol bacteria pellet and converting the carbon source to glycolic acid using the modified strain. The polyvinyl alcohol bacteria pellet and an auxiliary solution are mixed, and air is introduced while supplying the carbon source to the polyvinyl alcohol bacteria pellet. [Selection Diagram] Figure 8

Inventors

  • 袁 敬堯
  • 陳 仲裕
  • 李 奕成
  • 翁 梓桓
  • 簡 良榮

Assignees

  • 南亞塑膠工業股▲分▼有限公司

Dates

Publication Date
20260507
Application Date
20250212
Priority Date
20241230

Claims (10)

  1. To provide a modified strain containing the gene sequences of SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, To prepare polyvinyl alcohol bacteria pellets containing the modified strain, The polyvinyl alcohol bacteria pellet and the auxiliary solution are mixed, and ethylene glycol, which is a carbon source, is supplied to the polyvinyl alcohol bacteria pellet to convert the carbon source to glycolic acid using the modified strain. Includes, Air is introduced to the polyvinyl alcohol bacterial pellet while supplying the carbon source. A method for producing glycolic acid by an immobilization technique characterized by the above.
  2. The preparation of the polyvinyl alcohol bacterial pellets is A 1% to 20% polyvinyl alcohol solution is prepared, and the modified strain is added to the polyvinyl alcohol solution and mixed uniformly to obtain a polyvinyl alcohol bacterial solution. Prepare saturated boric acid solution and phosphoric acid solution, The polyvinyl alcohol bacterial solution is dropped into the saturated boric acid solution to obtain a semi-finished polyvinyl alcohol bacterial pellet product. The polyvinyl alcohol bacterial pellet semi-finished product is removed, drained, and placed in the phosphoric acid solution and left for a predetermined time to obtain the polyvinyl alcohol bacterial pellet. including, A method for producing glycolic acid by the immobilization technique described in claim 1.
  3. The preparation of the polyvinyl alcohol bacterial pellets further includes preparing 0.5% to 1.5% alginic acid. A method for producing glycolic acid by the immobilization technique described in claim 2.
  4. The aforementioned predetermined time is 30 minutes to 2 hours. A method for producing glycolic acid by the immobilization technique described in claim 2.
  5. The concentration of the saturated boric acid solution is 1% to 5%, and the concentration of the phosphoric acid solution is 0.1 M to 1 M. A method for producing glycolic acid by the immobilization technique described in claim 2.
  6. The airflow rate of the aforementioned air is 0.3 vvm to 1 vvm. A method for producing glycolic acid by the immobilization technique described in claim 1.
  7. The aforementioned modified strains are modified Escherichia coli, Gluconobacter sp . , Rhodococcus, or oxidizing bacteria. A method for producing glycolic acid by the immobilization technique described in claim 1.
  8. The aforementioned auxiliary solution is composed of yeast extract, peptone, sorbitol, ( NH₄ ) ₂SO₄ and MgSO₄ · 7H₂O . A method for producing glycolic acid by the immobilization technique described in claim 1.
  9. The GC% for SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3 are all between 50% and 60%. A method for producing glycolic acid by the immobilization technique described in claim 1.
  10. The modified strain has the ability to produce alcohol dehydrogenase, aldehyde dehydrogenase, and bacterial hemoglobin. A method for producing glycolic acid by the immobilization technique described in claim 1.

Description

This invention relates to a method for producing glycolic acid, and more particularly to a method for producing glycolic acid using immobilization technology. Glycolic acid (GA) is the smallest α-hydroxy acid, also known as hydroxyacetic acid or ethanolic acid. Glycolic acid contains both carboxyl and hydroxyl groups, possessing the dual properties of a carboxylic acid and an alcohol. Therefore, glycolic acid is easily decomposed and absorbed, possesses strong water solubility, and high permeability. Furthermore, glycolic acid and its polymers are highly biodegradable and biocompatible, decomposing within living organisms and being metabolized into water and carbon dioxide for excretion. For this reason, glycolic acid has a very wide range of applications. For example, glycolic acid and its polymers can be used in the release of peptide and protein-based pharmaceuticals, or as pharmaceutical intermediates in the production of menthol and quinine esters and the synthesis of other pharmaceuticals. Glycolic acid oligomers or derivatives are used as food additives, inhibiting the growth of harmful microorganisms through acidification. Conventionally, glycolic acid is produced primarily by chemical synthesis (e.g., cyanide of formaldehyde, chloroacetic acid hydrolysis, and oxonio of formaldehyde). However, these chemical synthesis methods have problems such as extreme reaction conditions, highly toxic raw materials, high costs, difficulty in subsequent separation, and environmental pollution. Therefore, with the spread of microbial enzyme-catalyzed methods and total biosynthesis, these are gradually becoming the mainstream methods for producing glycolic acid, replacing chemical synthesis. This is a flowchart of the method for converting a carbon source to glycolic acid according to the present invention.This is a schematic diagram of the plasmid according to the present invention.This is a schematic diagram showing a comparison between SEQ ID NO:1 according to the present invention and an existing EGADH gene sequence.Figures 4A and 4B are schematic diagrams showing a comparison between SEQ ID NO:2 according to the present invention and an existing EGAlDH gene sequence.This is a schematic diagram showing a comparison between SEQ ID NO:3 according to the present invention and an existing VHb gene sequence.These are bar graphs and curve graphs showing the amount of glycolic acid converted according to the first embodiment of the present invention.These are bar graphs and curve graphs showing the amount of glycolic acid converted according to the second embodiment of the present invention.This is a flowchart showing the method for producing glycolic acid using immobilization technology. The following describes embodiments of the "method for producing glycolic acid by immobilization technology" disclosed herein, using specific examples. Those skilled in the art will be able to understand the advantages and effects of the present invention from the disclosed content. The present invention can be implemented or applied through other different specific embodiments, and various detailed descriptions herein can be modified and altered in various ways based on different perspectives and applications, without departing from the spirit of the invention. Furthermore, it should be noted that the drawings of the present invention are for illustrative purposes only and are not based on actual dimensions. The following embodiments will describe the technical content of the present invention in more detail, but the disclosed content is not intended to limit the scope of protection of the present invention. In this specification, terms such as "first," "second," and "third" may be used to describe various elements or signals, but it should be understood that these elements or signals should not be limited by these terms. These terms are primarily used to distinguish one element from another, or one signal from another. Furthermore, the term "or" in this specification should be understood to include any one or more of the items listed in relation to it, depending on the actual context. The present invention will be explained with reference to Figures 1 and 2. The present invention provides a method for converting a carbon source to glycolic acid. This method includes step S10 of providing a plasmid containing the gene sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3; step S20 of introducing the plasmid into a bacterial strain to obtain a modified strain; and step S30 of supplying a carbon source to the modified strain and converting the carbon source to glycolic acid by the modified strain. More specifically, a step of synthesizing a DNA sequence may be further included before step S10. In the step of synthesizing a DNA sequence, a sequence having genetic coding recognition capabilities adapted to the target bacterial strain is artificially synthesized so that the target bacterial strain can recognize and produce the corres