KR-102963611-B1 - Method for preparing of unsaponifiable isolated from microalgae and unsaponifiable isolated from microalgae prepared by the method

Abstract

The present invention relates to a method for producing unsaponifiable matter derived from microalgae and unsaponifiable matter derived from microalgae produced thereby, comprising: (A) a step of mixing microalgae with a first solvent and performing a first extraction at 40 to 70 ℃; (B) a step of performing a second extraction of the first extract at 100 to 300 rpm in a shaking culture medium and then allowing it to stand to separate microalgae lipids through solid-liquid separation; and (C) a step of mixing microalgae lipids with a second solvent and performing a saponification reaction with an ultrasonic device to obtain unsaponifiable matter that has not been saponified; thereby, unsaponifiable matter derived from microalgae that can be used in compositions for osteoporosis, antifungal, anti-inflammatory, and antioxidant purposes can be obtained.

Inventors

• 김진우
• 염서희
• 김소희
• 박하영
• 강민호

Assignees

• 선문대학교 산학협력단

Dates

Publication Date

20260513

Application Date

20230504

Priority Date

20230125

Claims (6)

1. (A) A step of mixing microalgae and a first solvent and performing a first extraction at 40 to 70 ℃; (B) a step of separating microalgae lipids by solid-liquid separation after performing a second extraction of the above first extract in a shaking culture at 100 to 300 rpm and allowing it to stand; and (C) a step of mixing the microalgae lipid and the second solvent and performing a saponification reaction at 60 to 100°C for 50 to 150 minutes using an ultrasonic device with a frequency of 20 to 50 kHz and a power of 50 to 700 W, and then obtaining unsaponifiable material; comprising, A method for producing unsaponifiable matter derived from microalgae, characterized in that, in step (C) above, the second solvent is one selected from the group consisting of potassium hydroxide, sodium hydroxide, magnesium hydroxide, and calcium hydroxide; and a mixed solvent comprising an aqueous solution of a lower alcohol having 1 to 4 carbon atoms.
2. A method for producing unsaponifiable matter derived from microalgae according to claim 1, wherein in step (A), the first solvent is hexane, a lower alcohol having 1 to 4 carbon atoms, ethylene glycol, ethyl ether, or a mixture thereof.
3. delete
4. delete
5. In claim 1, the microalgae are Chlorella , Scenedesmus , Monoraphidium , Spirulina , Haematococcus pluvialis , Coscinodiscus granii, Amphora pediculus , Fragilaria oceanica , Coscinodiscus rothii , Amphora ovalis , Navicula sp ., Achnanthidium sp2 , Naviculara mosissinma , Alexandrium sp. cf. catenella , Actinocyclus normani ( A method for producing unsaponifiable matter derived from microalgae, characterized by being one or more selected from the group consisting of Actinocyclus normanii , Akashiwo sanguinea , Eucampia zodiacus , Ditylum brightwellii , Tetraselmis suecica , and Nitzschia sp3 .
6. Microalgae-derived unsaponifiable matter produced according to the manufacturing method of any one of paragraphs 1, 2, and 5.

Description

Method for preparing of unsaponifiable isolated from microalgae and unsaponifiable isolated from microalgae prepared by the method The present invention relates to a method for producing unsaponifiable matter derived from microalgae and unsaponifiable matter derived from microalgae produced according to this method. Algae are a group of organisms that collectively refer to photosynthetic organisms, with approximately 30,000 to over 1 million species reported to date. Among them, microalgae are single-celled microorganisms that perform photosynthesis and are gaining attention as raw materials capable of producing nutrients such as proteins, carbohydrates, lipids, and minerals, as well as physiologically active substances such as beta-glucan and astaxanthin. The aforementioned microalgae are autotrophic organisms that synthesize lipids, pigments, and proteins through photosynthesis. Since the content and composition of useful substances vary even within the same species depending on the culture environment, there is growing interest in increasing the production of physiologically active substances based on the growth characteristics of microalgae. In addition, since there are differences in cell growth and the production of intracellular lipids and pigments in microalgae depending on light intensity, temperature, and aeration, there is a trend of conducting in-depth research on culture factors and optimal culture processes to simultaneously improve cell mass and lipid content, which are primary metabolites, from microalgae. As the Food and Drug Administration (FDA) classifies microalgae such as Chlorella sp . and Spirulina sp . as safe materials that can be added to food, microalgae are currently being utilized in the food industry. Microalgae are gaining attention as a third-generation biomass. They have the advantage of high productivity and do not have the disadvantages of first and second-generation biomass, such as the need for arable land, the complexity of pretreatment processes, and low efficiency. As microalgae, which have a lipid content 50 to 100 times higher than conventional edible crops, are gaining attention in various fields, microalgae utilization technologies are evolving into cultivation, harvesting, lipid extraction, and biofuel conversion. Meanwhile, plant lipids are generally triglycerides that decompose into glycerol and alkali salts of fatty acids through a heating reaction in an alkaline phase. The alkali salts of the fatty acids are a type of saponified product, and the corresponding reaction is called the saponification reaction. Substances that are not saponified through the above saponification reaction are collectively referred to as unsaponifiable matter, and unsaponifiable matter contains various useful substances. Therefore, a method is required to reduce the impurity content and obtain a large amount of unsaponifiable matter with high purity. Figures 1a to 1c are graphs of microalgae lipids prepared according to Example 1 of the present invention measured by GC-MS. FIGS. 2a to 2d are graphs measuring unsaponifiable matter prepared according to Example 1 of the present invention. The present invention relates to a method for producing unsaponifiable matter derived from microalgae and unsaponifiable matter derived from microalgae produced according to this method. The present invention will be described in detail below. The method for preparing microalgae-derived unsaponifiable matter according to the present invention may comprise: (A) a step of mixing microalgae with a first solvent and performing a first extraction at 40 to 70 ℃; (B) a step of performing a second extraction of the first extract at 100 to 300 rpm in a shaking culture medium and then allowing it to stand to separate microalgae lipids through solid-liquid separation; and (C) a step of mixing the microalgae lipids with a second solvent and performing a saponification reaction with an ultrasonic device to obtain unsaponifiable matter. First, in step (A) above, a first extraction is performed by mixing microalgae and a first solvent at 40 to 70 ℃, and then in step (B) above, the first extract is secondly extracted at 100 to 300 rpm in a shaking culturer, and then the microalgae lipids are separated by solid-liquid separation by letting it stand. The above microalgae lipid is not particularly limited as long as the method for obtaining lipids is available, but preferably, it is obtained by first extracting using hexane, a lower alcohol having 1 to 4 carbon atoms, ethylene glycol, ethyl ether, or a mixed solvent thereof (first solvent), followed by second extraction in a shaking culture medium. By using the microalgae lipid obtained by the method for obtaining microalgae lipids according to the present invention, a large amount of unsaponifiable matter can be obtained thereafter. Examples of the above lower alcohols include aqueous solutions of methanol, ethanol, butanol, or propanol in an amount of 20 to 99 volume%