CN111184744B - Hepu pearl extract with antibacterial effect and preparation method thereof - Google Patents
Hepu pearl extract with antibacterial effect and preparation method thereof Download PDFInfo
- Publication number
- CN111184744B CN111184744B CN202010049744.8A CN202010049744A CN111184744B CN 111184744 B CN111184744 B CN 111184744B CN 202010049744 A CN202010049744 A CN 202010049744A CN 111184744 B CN111184744 B CN 111184744B
- Authority
- CN
- China
- Prior art keywords
- extract
- hepu
- pearl powder
- hepu pearl
- acid solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000000284 extract Substances 0.000 title claims abstract description 100
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 23
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- 239000000843 powder Substances 0.000 claims abstract description 52
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 12
- 229920001184 polypeptide Polymers 0.000 claims abstract description 10
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 10
- 238000000605 extraction Methods 0.000 claims abstract description 8
- 238000000034 method Methods 0.000 claims description 31
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 27
- 241000191967 Staphylococcus aureus Species 0.000 claims description 23
- 238000003756 stirring Methods 0.000 claims description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 239000006228 supernatant Substances 0.000 claims description 14
- 239000002253 acid Substances 0.000 claims description 12
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 12
- 241000588724 Escherichia coli Species 0.000 claims description 11
- 241000589517 Pseudomonas aeruginosa Species 0.000 claims description 11
- 238000009835 boiling Methods 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 9
- 230000008569 process Effects 0.000 claims description 9
- 239000000047 product Substances 0.000 claims description 8
- 230000002401 inhibitory effect Effects 0.000 claims description 7
- 239000004310 lactic acid Substances 0.000 claims description 6
- 235000014655 lactic acid Nutrition 0.000 claims description 6
- 230000000845 anti-microbial effect Effects 0.000 claims description 5
- 239000002994 raw material Substances 0.000 claims description 4
- 150000003384 small molecules Chemical class 0.000 claims description 4
- 238000002604 ultrasonography Methods 0.000 claims description 4
- 238000009777 vacuum freeze-drying Methods 0.000 claims description 4
- 241000293871 Salmonella enterica subsp. enterica serovar Typhi Species 0.000 claims description 2
- 230000002147 killing effect Effects 0.000 claims description 2
- 239000003755 preservative agent Substances 0.000 claims description 2
- 230000002335 preservative effect Effects 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- 238000002525 ultrasonication Methods 0.000 claims description 2
- 230000003385 bacteriostatic effect Effects 0.000 abstract description 25
- 102000004142 Trypsin Human genes 0.000 abstract description 7
- 108090000631 Trypsin Proteins 0.000 abstract description 7
- 239000012588 trypsin Substances 0.000 abstract description 7
- 239000003814 drug Substances 0.000 abstract description 5
- 229910021645 metal ion Inorganic materials 0.000 abstract description 5
- 238000010257 thawing Methods 0.000 abstract description 4
- 229940079593 drug Drugs 0.000 abstract description 3
- 239000000654 additive Substances 0.000 abstract description 2
- 238000011161 development Methods 0.000 abstract description 2
- 230000007613 environmental effect Effects 0.000 abstract description 2
- 230000002421 anti-septic effect Effects 0.000 abstract 1
- 235000013376 functional food Nutrition 0.000 abstract 1
- 239000011049 pearl Substances 0.000 description 111
- 239000000243 solution Substances 0.000 description 26
- 230000000694 effects Effects 0.000 description 20
- 238000002474 experimental method Methods 0.000 description 18
- 238000011160 research Methods 0.000 description 10
- 241000193830 Bacillus <bacterium> Species 0.000 description 9
- 208000037386 Typhoid Diseases 0.000 description 9
- 230000005764 inhibitory process Effects 0.000 description 9
- 201000008297 typhoid fever Diseases 0.000 description 9
- 239000013543 active substance Substances 0.000 description 8
- 238000012360 testing method Methods 0.000 description 8
- 102000004190 Enzymes Human genes 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 6
- 229940088598 enzyme Drugs 0.000 description 6
- 239000013642 negative control Substances 0.000 description 6
- 102000057297 Pepsin A Human genes 0.000 description 5
- 108090000284 Pepsin A Proteins 0.000 description 5
- 239000004365 Protease Substances 0.000 description 5
- 229940111202 pepsin Drugs 0.000 description 5
- 239000008223 sterile water Substances 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 238000005303 weighing Methods 0.000 description 4
- 102000035195 Peptidases Human genes 0.000 description 3
- 108091005804 Peptidases Proteins 0.000 description 3
- 241000490567 Pinctada Species 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 239000004480 active ingredient Substances 0.000 description 3
- 239000011575 calcium Chemical group 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 235000019419 proteases Nutrition 0.000 description 3
- 241000250967 Branchia Species 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical group [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- 206010010904 Convulsion Diseases 0.000 description 2
- 206010012735 Diarrhoea Diseases 0.000 description 2
- 241000237852 Mollusca Species 0.000 description 2
- 241000475481 Nebula Species 0.000 description 2
- 108090000526 Papain Proteins 0.000 description 2
- 241000490568 Pinctada fucata Species 0.000 description 2
- 241001212699 Pinctada martensii Species 0.000 description 2
- 241001464055 Pteriidae Species 0.000 description 2
- 230000000941 anti-staphylcoccal effect Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 229960005069 calcium Drugs 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 230000036461 convulsion Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 201000004792 malaria Diseases 0.000 description 2
- 229940055729 papain Drugs 0.000 description 2
- 235000019834 papain Nutrition 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 230000002087 whitening effect Effects 0.000 description 2
- 208000002874 Acne Vulgaris Diseases 0.000 description 1
- 206010014970 Ephelides Diseases 0.000 description 1
- 206010062717 Increased upper airway secretion Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 239000012880 LB liquid culture medium Substances 0.000 description 1
- 208000003351 Melanosis Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010059013 Nocturnal emission Diseases 0.000 description 1
- 208000001132 Osteoporosis Diseases 0.000 description 1
- 206010033557 Palpitations Diseases 0.000 description 1
- 229930003270 Vitamin B Chemical group 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 206010000496 acne Diseases 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 235000001465 calcium Nutrition 0.000 description 1
- MKJXYGKVIBWPFZ-UHFFFAOYSA-L calcium lactate Chemical compound [Ca+2].CC(O)C([O-])=O.CC(O)C([O-])=O MKJXYGKVIBWPFZ-UHFFFAOYSA-L 0.000 description 1
- 239000001527 calcium lactate Substances 0.000 description 1
- 229960002401 calcium lactate Drugs 0.000 description 1
- 235000011086 calcium lactate Nutrition 0.000 description 1
- 230000001914 calming effect Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 235000021466 carotenoid Nutrition 0.000 description 1
- 150000001747 carotenoids Chemical group 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 238000010227 cup method (microbiological evaluation) Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 230000004438 eyesight Effects 0.000 description 1
- 230000001815 facial effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000003179 granulation Effects 0.000 description 1
- 238000005469 granulation Methods 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 108010059345 keratinase Proteins 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 239000002398 materia medica Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 230000024121 nodulation Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000011164 ossification Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 208000026435 phlegm Diseases 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 150000004032 porphyrins Chemical class 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 235000015170 shellfish Nutrition 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011720 vitamin B Chemical group 0.000 description 1
- 235000019156 vitamin B Nutrition 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/618—Molluscs, e.g. fresh-water molluscs, oysters, clams, squids, octopus, cuttlefish, snails or slugs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Communicable Diseases (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Zoology (AREA)
- Marine Sciences & Fisheries (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Organic Chemistry (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention provides a Hepu pearl powder extract containing small molecular active polypeptide, a preparation method and application thereof. Compared with zymolyte, the Hepu pearl extract prepared by the invention has strong bacteriostatic activity, has stable bacteriostatic activity under acid-base conditions, and has better stability to temperature, repeated freeze thawing, ultraviolet rays, metal ions and trypsin. In addition, the invention also has the advantages of simple extraction method, simple equipment, energy saving, environmental protection and high extraction rate, and has good development and application prospects in the aspects of medicines, high-grade health products, functional foods, antibacterial and antiseptic additives and the like.
Description
Technical Field
The invention belongs to the technical field of active ingredient extraction, and particularly relates to a small molecular active polypeptide substance with bacteriostatic activity extracted from Hepu pearl powder, a preparation method and an antibacterial application thereof.
Background
Pinctada martensii (Pinctada martensii), also known as Pinctada fucata (Pinctada fucata), belongs to the phylum Mollusca (Mollusca), the class Lamellia branchia (Lamelli branchia), the order Pinctada (Pteriidaea), the family Pteriidae (Pteriidae), the genus Pinctada (Pinctada), and is the main shellfish for producing seawater pearls in south China, mainly distributed in Guangxi, guangdong and Hainan in China. Hepu pearl is produced in Hepu in North sea of Guangxi, and is a geographical sign product in China. Besides being used as a noble and elegant ornament, hepu pearl is also a traditional and noble medicinal material, and has a medicinal history for thousands of years in China. Many traditional Chinese medical classics have definite records on the property and taste of Hepu pearls: the record of Shen nong Ben Cao Jing in Qin Han period states that pearl powder can remove cold and heat, damp malaria \8230andwet malaria \8230, and after long-term administration, it can strengthen bone joint, kill evil ghost and prolong life; the medical book of three kingdoms (famous medical records) has listed pearls as important medicinal materials and clarified the drug effect; the pearl recorded in the book of the collection and the comments of the materia medica channel of the south and north dynasties of ceramic and Hongjing has the functions of treating nebula and stopping diarrhea; the record of Ben Cao gang mu written by Ming dynasty Li Shizhen: pearl is salty in taste, sweet, cold and nontoxic. Relieving palpitation and nodulation. The coating is smooth and has good color. The paste is applied to hands and feet, peeled, retrogradable, and has the effects of eliminating phlegm, removing facial speckle and stopping diarrhea. Can remove infantile convulsion and tranquilize. Stopping nocturnal emission, eliminating acne and healing toxin. The modern book of pharmacopoeia of the people's republic of China and Chinese medicine dictionary records: the pearl has the effects of calming nerves and arresting convulsion, improving eyesight and removing nebula, detoxifying and promoting granulation and moistening skin.
The pearl powder is from ancient homology of medicine and food, and is popular as a pure natural beauty-maintaining, health-preserving and health-care product for thousands of years. For example, CN110240627A discloses a method for extracting small molecule active peptide from pearl powder by using complex enzyme, which comprises the following steps: adding pearl powder into acid solution for acidolysis, and collecting supernatant and first insoluble protein; soaking the first insoluble protein in water, and centrifuging to obtain a second supernatant and a second insoluble protein; combining the first supernatant and the second supernatant; vacuum concentrating the combined first supernatant and second supernatant to separate out calcium lactate, filtering, mixing with the second insoluble protein, and homogenizing to obtain homogenate; carrying out enzymolysis on the homogenate under a weak acid condition by using a compound protease solution to obtain an enzymolysis solution, wherein the compound protease solution comprises a trypsin solution, a papain solution and a keratinase solution; decolorizing and sterilizing the enzymolysis solution, and making into small molecule active peptide with whitening effect in liquid or powder form.
The current research shows that the main active substances of the Hepu pearl comprise trace elements, porphyrin and metalloporphyrin compounds, calcium, carotenoid, vitamin B group, small molecule active polypeptide, polysaccharide, amino acid and other important components. The Hepu pearl powder has the characteristics of obvious effect, no toxic or side effect and the like, and is often applied to the compatibility of various formulas and health-care products to play the effect. The active substance identification and pharmacological action of Hepu pearl powder are the research directions of many researchers. Modern researches show that the pearl powder has various effects including multiple biological activities of regulating the immune function of the body, resisting inflammation, fatigue, aging, oxidation, whitening, removing freckles, supplementing calcium, resisting cancer, reducing blood sugar, reducing blood fat, resisting osteoporosis, promoting osteogenesis and the like, but relatively few researches on the antibacterial aspect of Hepu pearl powder are carried out.
Disclosure of Invention
In view of the defects of the prior art, the invention aims to provide a Hepu pearl powder extract containing small molecular active polypeptide, a preparation method and application thereof, wherein the extract can obviously inhibit the growth of staphylococcus aureus, escherichia coli, typhoid bacillus and pseudomonas aeruginosa and can be used for medicines.
In order to achieve the technical purpose, the inventor combines the research experience of the Hepu pearl for many years, and through a large number of experimental researches and continuous exploration, and finally extracts the Hepu pearl powder extract with remarkable bacteriostatic activity by adopting an intermittent ultrasonic wave and acidolysis method.
The research of the invention obtains the subsidies of Guangxi natural science fund youth scientific fund (No. 2018GXNSFBA281100), guangxi university middle-aged and youth teacher scientific research basic capability improvement project fund (No. 2019KY0327) and Guangxi traditional Chinese medicine university doctor scientific research starting fund (No. 2017BS006) in 2017.
Specifically, the technical scheme of the invention is summarized as follows:
a Hepu pearl powder extract containing small molecular active polypeptide is prepared from Hepu pearl powder as raw material by intermittent ultrasonic wave with acidolysis extraction and centrifugal separation, wherein the acid solution used for acidolysis is 1-10% acetic acid solution or lactic acid solution, the intermittent ultrasonic power is 200-300W, the working time is 4-6s, the intermittent time is 4-6s, and the whole course is 25-35min.
A preparation method of Hepu pearl powder extract containing small molecular active polypeptide comprises the following steps:
(1) Taking Hepu pearl powder, adding 1-10% acid solution which is acetic acid solution or lactic acid solution, and carrying out ultrasonic crushing at room temperature, wherein the ultrasonic conditions are as follows: the power is 200-300W, the working time is 4-6s, the intermittence time is 4-6s, and the whole process lasts 25-35min;
(2) Continuously stirring for 4-8h after the ultrasonic disruption is finished, adjusting the pH value to 7.0-7.4, transferring to a constant temperature environment of 37-42 ℃, stirring overnight, keeping in a boiling water bath for 10-20min after the stirring is finished, centrifuging by adopting a refrigerated centrifuge, taking the supernatant, adjusting the pH value to 7.0-7.4, and performing vacuum freeze drying to obtain the Hepu pearl powder extract.
Further preferably, in the preparation method of the extract of the Hepu pearl powder, the acid solution is 4-6% acetic acid solution, and the ratio of the material to the liquid of the Hepu pearl powder to the acid solution is 1: (30-60).
Further preferably, the preparation method of the extract of Hepu pearl powder as described above, wherein the ultrasonic conditions are as follows: 250W of power, 5sec of working time, 5sec of pause time and 30min of whole course.
Further preferably, the method for preparing the extract of Hepu pearl powder as described above, wherein the stirring speed in step (2) is 500-800rpm.
Further preferably, the method for preparing the extract of Hepu pearl powder as described above, wherein after the ultrasonication is completed, the stirring is continued for 6h, then the pH is adjusted to =7.0, and the mixture is transferred to a constant temperature environment of 40 ℃ and stirred overnight, and then kept in a boiling water bath for 15min after the completion.
Further preferably, the preparation method of the extract of Hepu pearl powder as described above, wherein the technical parameters of the centrifugal of the refrigerated centrifuge are as follows: centrifuge at 5000rpm for 15min at 4 ℃.
In addition, the invention takes Hepu pearl powder as raw material, extracts effective active substances in the raw material, adopts methods such as an inhibition zone, minimum inhibition concentration, a liquid growth curve and the like to evaluate the inhibition effect of the Hepu pearl extract, discusses the stability of the inhibition activity, compares the inhibition activity with the Hepu pearl hydrolysate prepared by an enzymolysis method, and provides theoretical basis for developing natural bacteriostat and effective utilization of active ingredients of the Hepu pearl.
Based on the research result, the invention also provides the application of the extract of the Hepu pearl powder containing the small molecular active polypeptide in preparing antibacterial products. The antibacterial product has inhibitory or killing activity on staphylococcus aureus, escherichia coli, typhoid bacillus and pseudomonas aeruginosa. Further preferably, the antimicrobial product is a pharmaceutical or preservative.
Compared with the prior art, the Hepu pearl powder extract and the preparation method thereof provided by the invention have the following advantages and progresses:
(1) The Hepu pearl extract has very obvious inhibitory activity on the growth of staphylococcus aureus, escherichia coli, typhoid bacillus and pseudomonas aeruginosa, and the activity is obviously higher than that of the extract prepared by an enzymolysis method.
(2) The active ingredients in the Hepu pearl extract have stable bacteriostatic activity under acid-base conditions, have good stability on temperature, repeated freeze thawing and ultraviolet rays, and have good stability on metal ions Na + 、K + 、Ca 2+ And Mg 2+ Also has better stability and simultaneously has better stability to trypsin.
(3) The invention also has the advantages of simple extraction method, simple equipment, energy saving, environmental protection and high extraction rate which reaches 83.2 percent, and has good development and application prospects in the aspects of medicines, antibacterial and anticorrosive additives and the like.
Drawings
FIG. 1 is a photograph of a lyophilized powder of Hepu pearl extract according to the present invention;
FIG. 2 is the bacteriostatic activity of the extract of Hepu pearl prepared in example 1 against different species, wherein A: (ii) staphylococcus aureus; b: escherichia coli; c: typhoid bacillus; d: pseudomonas aeruginosa;
FIG. 3 shows the results of the determination of the minimum inhibitory concentration of different species from the Hepu pearl prepared in example 1, wherein A: (ii) staphylococcus aureus; b: e.coli; c: typhoid bacillus; d: pseudomonas aeruginosa;
FIG. 4 is a graph of the effect of Hepu pearl extract prepared in example 1 on the growth of Staphylococcus aureus;
FIG. 5 is a graph of the effect of temperature (A) and repeated freeze-thaw cycles (B) on the anti-Staphylococcus aureus activity of the Hepu pearl extract prepared in example 1;
FIG. 6 is a graph showing the effect of pH (A) and UV light (B) on the antibacterial activity of the extract of Hepu pearl prepared in example 1;
fig. 7 is a graph showing the effect of metal ions on the bacteriostatic activity of the extract of Hepu pearl prepared in example 1, wherein A: naCl; b: KCl; c: caCl 2 ；D：MgCl 2 ；
FIG. 8 shows the effect of trypsin and pepsin on the antibacterial activity of the extract of Hepu pearl prepared in example 1.
Detailed Description
The invention will be further described with reference to examples and drawings, the advantages and features of which will become more apparent as the description proceeds. It should be understood that the illustrated embodiments are exemplary only, and are not intended to limit the scope of the present invention in any way. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit of the invention, and it is intended to cover all such changes and modifications as fall within the scope of the invention.
The experimental procedures in the following examples are all conventional ones unless otherwise specified. The Hepu pearl powder in the following examples was purchased from Baozhilin ocean technologies, inc., north sea, guangxi.
Example 1: preparation of Hepu pearl extract (ultrasound + acetic acid)
Accurately weighing 5.0g of Hepu pearl powder, adding 200mL of 5% acetic acid solution, uniformly stirring, and carrying out ultrasonic crushing at room temperature under the crushing conditions: the power is 250W, the working time is 5sec, the intermittence time is 5sec, and the whole process is 30min; then, the mixture is magnetically stirred at 600rpm for 6 hours at room temperature, the pH value is adjusted to 7.0, the mixture is transferred to a thermostat with the temperature of 40 ℃ and magnetically stirred at 600rpm for overnight, the mixture is kept in boiling water for 15 minutes after the stirring is finished, the mixture is cooled and then centrifuged at 4 ℃ and 000rpm for 15 minutes, the supernatant is taken out, the pH value is adjusted to 7.0, the mixture is freeze-dried into powder in vacuum, and the powder is stored at room temperature (see figure 1).
Example 2: preparation of Hepu pearl extract (ultrasound + lactic acid)
Accurately weighing 5.0g of Hepu pearl powder, adding 200mL of 5% lactic acid solution, stirring uniformly, carrying out ultrasonic crushing at room temperature, and carrying out crushing conditions: the power is 250W, the working time is 5sec, the intermittence time is 5sec, and the whole process is 30min; then continuing to magnetically stir at 600rpm for 6h at room temperature, adjusting pH to 7.0, transferring to a thermostat at 40 ℃ and magnetically stirring at 600rpm overnight, keeping in boiling water for 15min after finishing, cooling, centrifuging at 4 ℃ for 15min at 5,000rpm, taking supernatant, adjusting pH to 7.0, freeze-drying in vacuum to obtain powder, and storing at room temperature.
Comparative example 1: preparation of Hepu pearl extract (ultrasound + hydrochloric acid)
Accurately weighing 5.0g of Hepu pearl powder, adding 200mL of 5% hydrochloric acid solution, uniformly stirring, and carrying out ultrasonic crushing at room temperature under the crushing conditions: the power is 250W, the working time is 5sec, the intermittence time is 5sec, and the whole process is 30min; then continuing to magnetically stir at 600rpm for 6h at room temperature, adjusting pH to 7.0, transferring to a thermostat at 40 ℃ and magnetically stirring at 600rpm overnight, keeping in boiling water for 15min after finishing, cooling, centrifuging at 4 ℃ for 15min at 5,000rpm, taking supernatant, adjusting pH to 7.0, freeze-drying in vacuum to obtain powder, and storing at room temperature.
Comparative example 2: preparation of Hepu pearl zymolyte (enzymolysis extraction)
Accurately weighing 5.0g of Hepu pearl powder, adding 200mL of 5% acetic acid solution, stirring uniformly, carrying out ultrasonic crushing at room temperature, wherein the crushing conditions are as follows: the power is 250W, the working time is 5sec, the pause time is 5sec, and the whole process lasts 30min; then, the magnetic stirring is carried out at the room temperature at the speed of 600rpm for 6 hours, the pH is adjusted to 7.0, 0.25g of papain is added according to the proportion of 5 percent (the weight percent of the enzyme is generally 2 to 8 percent of that of the pearl powder). Hydrolyzing at 40-50 deg.C overnight, keeping in boiling water for 15min to inactivate enzyme after hydrolysis, cooling, centrifuging at 4 deg.C and 5,000rpm for 15min, collecting supernatant, adjusting pH to 7.0, vacuum freeze drying to obtain powder, and storing at room temperature.
Example 3: in vitro antibacterial activity test of Hepu pearl extract
An antibacterial activity experiment is carried out by adopting an oxford cup method: respectively picking single colonies of staphylococcus aureus, escherichia coli, typhoid bacillus and pseudomonas aeruginosa from an LB solid plate, inoculating the single colonies into 5mL of fresh LB liquid culture medium, carrying out shaking culture at 37 ℃ and 180r/min for 4h to a logarithmic phase of growth, and diluting to 1 × 10 6 CFU·mL -1 . Transferring 200 μ L of the bacterial suspension by pipette, uniformly coating on LB solid plate, placing sterilized and dried Oxford cup on the surface of the plate with sterile forceps, gently pressing and fixing, sucking 200 μ L of 240 mg/mL -1 The Hepu pearl extract (pH = 7.0) is added into an Oxford cup, a negative control group is 200 mu L of sterile water, each group is provided with three parallel experiments, after diffusion is carried out for 3h at 4 ℃, the negative control group is placed in an incubator at 37 ℃, the negative control group is cultured for 18-24h to observe the formation of a bacteriostatic circle, and the size of the bacteriostatic circle is measured by using a vernier caliper.
The experimental results of the inhibition zones for different strains are shown in fig. 2 and table 1, and the extract of the Hepu pearl prepared in the embodiments 1 and 2 of the invention has significant inhibition effect on staphylococcus aureus, escherichia coli, typhoid bacillus and pseudomonas aeruginosa. Comparative example 1 has no obvious bacteriostatic activity on escherichia coli, has certain bacteriostatic activity on staphylococcus aureus, typhoid bacillus and pseudomonas aeruginosa, but has lower sensitivity. Comparative example 2 has certain bacteriostatic activity on staphylococcus aureus, escherichia coli, typhoid bacillus and pseudomonas aeruginosa, but the sensitivity is lower.
TABLE 1 comparison of the zone of inhibition diameters (mm) of Hepu pearl extract for different strains
Example 4: MIC (minimum inhibitory concentration) determination experiment of Hepu pearl extract on different strains
The minimum inhibitory concentration of the Hepu pearl extract is measured by a two-fold gradient dilution method. The method is the same as the above method for preparing 1 × 10 strains of different strains 6 CFU·mL -1 And (4) bacterial suspension. MIC assays were performed using sterile 96-well plates, 100. Mu.L of diluted bacterial suspension was added to each well, followed by 100. Mu.LExtracts of Hepu pearl diluted in sterile water at different concentrations, each group of concentrations was as follows: 240.00, 120.00, 60.00, 30.00, 15.00, 7.50, 3.75, 1.88, 0.94, 0.47, and 0.24 mg-mL -1 The negative control group is sterile water, LB culture solution without adding bacteria solution and Hepu pearl extract is used as blank control, each group has 3 parallel holes, and is cultured in 37 deg.C incubator for 24 hr, and then OD is detected by enzyme labeling instrument 600 And calculating MIC of the Hepu pearl extract to different strains.
The extract of Hepu pearl prepared in the embodiment 1 of the present invention has MIC values of 0.94, 15.0 and 15.0mg/mL (see FIG. 3) for Staphylococcus aureus, escherichia coli, salmonella typhi and Pseudomonas aeruginosa, as measured by Minimum Inhibitory Concentration (MIC).
Example 5: experiment on influence of Hepu pearl extract on growth of staphylococcus aureus
Staphylococcus aureus concentration of 1X 10 prepared by the same method as above 6 CFU·mL -1 The bacterial suspension of (4). The negative control group was sterile water, and the experimental groups were 0.5 × MIC, 1 × MIC, 2 × MIC, 4 × MIC, 8 × MIC, 16 × MIC, 32 × MIC, 64 × MIC, and 128 × MIC, and were quickly mixed and incubated at 37 ℃ for static culture. Experiment time points of the Hepu pearl extract on the growth influence of the Hepu pearl extract include 18 time points of 0, 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51h and the like, a bacterium solution and LB culture solution of the Hepu pearl extract are not added as blank controls, each group is provided with three parallel experiments, and the influence of the Hepu pearl extract prepared in example 1 on the growth of the Staphylococcus aureus is detected by a 96-well method.
The test result in fig. 4 shows that when the extract of the Hepu pearl with the concentration of 0.5 × MIC is added into the LB medium, the growth of the Staphylococcus aureus is basically not influenced, the inhibition effect on the growth of the Staphylococcus aureus is more obvious along with the increase of the concentration of the extract, the growth of the Staphylococcus aureus is obviously inhibited when the concentration of the added extract of the Hepu pearl is more than or equal to 2 × MIC, and the delay period is obviously prolonged compared with the negative control.
Example 5: influence of temperature and repeated freeze thawing on antibacterial activity of Hepu pearl extract
Using 7.5 mg. ML -1 Experiments are carried out on the Hepu pearl extract (prepared in example 1) with the concentration, the Hepu pearl extract is treated at-80, -20, 4, 25, 37, 60, 80 and 100 ℃ for 30min respectively, staphylococcus aureus is used as indicator bacteria, three parallel experiments are set, and 96-hole methods are adopted to detect the influence of different temperatures on the bacteriostatic activity of the Hepu pearl extract respectively.
As can be seen from the test results in FIG. 5A, the Hepu pearl extract has good bacteriostatic activity against Staphylococcus aureus at-80 deg.C to 100 deg.C, and slightly fluctuates at 100 deg.C.
The use amount was 7.5 mg. ML -1 The Hepu pearl extract (prepared in example 1) was subjected to an experiment in which it was frozen at-20 deg.C, then rapidly thawed at room temperature with freeze-thaw times of 1, 2, 3, 4, 5 and 6, respectively, and a Hepu pearl extract that was not freeze-thawed was used as a control, three parallel experiments were set, and the effects of repeated freeze-thaw on the bacteriostatic activity of the Hepu pearl extract were respectively detected by 96-well method.
As can be seen from the test results in fig. 5B, repeated freezing and thawing did not affect the bacteriostatic activity of the extract. In conclusion, the antibacterial active substance in the Hepu pearl extract has good thermal stability.
Example 6: influence of pH value on antibacterial activity of Hepu pearl extract
120 mg. About.mL -1 The extract of Hepu pearl (prepared in example 1) was mixed with sterile water at pH 3.0, 5.0, 7.0, 9.0 and 11.0, respectively, at a ratio of 1 -1 The Hepu pearl extract is subjected to an experiment, and the influence of different pH values on the bacteriostatic activity of the Hepu pearl extract is analyzed by a 96-hole method.
As can be seen from the test results in fig. 6A, after the extract of the philips pearl is treated under acidic, neutral and alkaline conditions, although its bacteriostatic activity slightly fluctuates at pH 11.0, it shows a strong bacteriostatic activity with good stability as a whole, which indicates that the active substance in the extract of the philips pearl of the present invention is resistant to acid and alkali, can still maintain a stable structure, is not easily decomposed, and further exerts its function.
Example 7: influence of ultraviolet irradiation on bacteriostatic activity of Hepu pearl extract
The use amount was 7.5 mg. ML -1 The Hepu pearl extract (prepared in example 1) is subjected to an experiment, the Hepu pearl extract is placed under ultraviolet light in an open manner, and is respectively irradiated for 0, 5, 10, 15, 20, 25 and 30min, staphylococcus aureus is used as an indicator bacterium, three parallel experiments are set, and the influence of ultraviolet light on the bacteriostatic activity of the Hepu pearl extract is detected by adopting a 96-hole method.
The test result of fig. 6B shows that the anti-bacterial activity of the extract of the philips pearl is not substantially affected by the ultraviolet treatment, and the stable and good anti-bacterial activity is maintained, which indicates that the active substance in the extract of the philips pearl of the present invention is stable, can resist the damage of ultraviolet, and can ensure that the extract of the philips pearl can better exert the anti-bacterial activity in the natural environment.
Example 8: influence of metal ions on bacteriostatic activity of Hepu pearl extract
The use amount was 7.5 mg. ML -1 The Hepu pearl extract (prepared in example 1) of (1) was used in an experiment of 15 mg/mL -1 The extract of Hepu pearl is mixed with different concentrations of 0, 25, 50, 100 and 200 mmoL.L -1 NaCl, KCl, caCl 2 And MgCl 2 The solutions are mixed in equal volume, three parallel experiments are carried out, the mixture is placed at 37 ℃ for treatment for 1h, and then a 96-hole method is adopted to analyze the influence of different ionic strengths on the antibacterial activity of the Hepu pearl extract on staphylococcus aureus.
As shown in the test results of FIG. 7, the antibacterial activity of the Hepu pearl extract was not substantially affected by the treatment with different metal ions, although Mg 2+ Has slight influence on its bacteriostatic activity, and may be Mg 2+ Can be combined with active substances in the extract to have certain influence on the structure, so that the antibacterial activity of the extract is slightly fluctuated, but the antibacterial activity is kept stable overall.
Example 9: influence of trypsin and pepsin on bacteriostatic activity of Hepu pearl extract
Trypsin was added to 30 mg.mL at pH 7.0 -1 In the Hepu pearl extract (prepared in example 1), pepsin was added at 30 mg/mL of pH 2.0 -1 The final enzyme concentration in the Hepu pearl extract is 1.0 mg/mL -1 Treating at 37 deg.C for 1 hr, treating at 100 deg.C for 5min, terminating enzyme reaction, centrifuging at room temperature of 12,000rpm for 5min, collecting supernatant, adjusting pH to 7.0, filtering for sterilization, and using 7.5 mg/mL -1 Experiments are carried out on the Hepu pearl extract with the concentration, the Hepu pearl extract without enzyme treatment is used as a reference, three parallel experiments are set, and the 96-hole method is adopted to analyze the influence of protease on the bacteriostatic activity of the Hepu pearl extract.
The test results in fig. 8 show that, compared with the untreated group, the antimicrobial activity of the extract of the symphyseal pearl treated by trypsin slightly fluctuates, but the significant antimicrobial activity is retained, and the extract treated by pepsin no longer has an antimicrobial effect on staphylococcus aureus, which indicates that pepsin can significantly degrade active substances in the extract of the symphyseal pearl, so that the anti-staphylococcus aureus activity of the extract disappears.
Claims (9)
1. The extract of Hepu pearl powder containing small molecular active polypeptide is characterized in that the extract of Hepu pearl powder is obtained by taking the Hepu pearl powder as a raw material, adopting intermittent ultrasonic wave to carry out acidolysis extraction and centrifugal separation, and the preparation method of the extract of Hepu pearl powder comprises the following steps:
(1) Taking Hepu pearl powder, adding 1-10% acid solution, wherein the acid solution is acetic acid solution or lactic acid solution, and ultrasonically crushing at room temperature, and the ultrasonic conditions are as follows: the power is 200-300W, the working time is 4-6s, the intermittence time is 4-6s, and the whole process lasts 25-35min;
(2) Continuously stirring for 4-8h after the ultrasonic disruption is finished, adjusting the pH value to 7.0-7.4, transferring to a constant temperature environment of 37-42 ℃, stirring overnight, keeping in a boiling water bath for 10-20min after the stirring is finished, centrifuging by adopting a refrigerated centrifuge, taking the supernatant, adjusting the pH value to 7.0-7.4, and performing vacuum freeze drying to obtain the Hepu pearl powder extract.
2. A preparation method of a Hepu pearl powder extract containing small molecular active polypeptide is characterized by comprising the following steps:
(1) Taking Hepu pearl powder, adding 1-10% acid solution which is acetic acid solution or lactic acid solution, and carrying out ultrasonic crushing at room temperature, wherein the ultrasonic conditions are as follows: the power is 200-300W, the working time is 4-6s, the intermittence time is 4-6s, and the whole process lasts 25-35min;
(2) Continuously stirring for 4-8h after the ultrasonic disruption is finished, adjusting the pH value to 7.0-7.4, transferring to a constant temperature environment of 37-42 ℃, stirring overnight, keeping in a boiling water bath for 10-20min after the stirring is finished, centrifuging by adopting a refrigerated centrifuge, taking the supernatant, adjusting the pH value to 7.0-7.4, and performing vacuum freeze drying to obtain the Hepu pearl powder extract.
3. The method of claim 2, wherein the acid solution is 4-6% acetic acid solution, and the ratio of the Hepu pearl powder to the acid solution is 1: (30-60).
4. The method for preparing Hepu pearl powder extract of claim 2, wherein the ultrasound conditions are as follows: 250W of power, 5s of working time, 5s of pause time and 30min of the whole process.
5. The method for preparing Hepu pearl powder extract of claim 2, wherein the stirring speed in step (2) is 500-800rpm.
6. The method for preparing Hepu pearl powder extract of claim 2, wherein the ultrasonication is performed, the stirring is continued for 6h, then the pH =7.0 is adjusted, the mixture is transferred to a constant temperature environment of 40 ℃ and stirred overnight, and then the mixture is kept in a boiling water bath for 15min after the completion.
7. The method of claim 2, wherein the technical parameters of the centrifugal process of the refrigerated centrifuge are as follows: centrifuge at 5000rpm for 15min at 4 ℃.
8. The use of extract of Hepu pearl powder containing small molecule active polypeptide of claim 1 for preparing antibacterial products with inhibitory or killing activity against Staphylococcus aureus, escherichia coli, salmonella typhi, and Pseudomonas aeruginosa.
9. Use according to claim 8, wherein the antimicrobial product is a pharmaceutical or preservative.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010049744.8A CN111184744B (en) | 2020-01-16 | 2020-01-16 | Hepu pearl extract with antibacterial effect and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010049744.8A CN111184744B (en) | 2020-01-16 | 2020-01-16 | Hepu pearl extract with antibacterial effect and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111184744A CN111184744A (en) | 2020-05-22 |
| CN111184744B true CN111184744B (en) | 2022-11-22 |
Family
ID=70684608
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010049744.8A Active CN111184744B (en) | 2020-01-16 | 2020-01-16 | Hepu pearl extract with antibacterial effect and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111184744B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016112722A1 (en) * | 2015-01-15 | 2016-07-21 | 欧诗漫生物股份有限公司 | Preparation method for conchiolin, and water-soluble conchiolin and acid-soluble conchiolin prepared by using method |
| CN106538991A (en) * | 2016-11-01 | 2017-03-29 | 福建农林大学 | A kind of method that ultrasonic acidolysis pre-treat biological prepares kelp blood pressure product |
| CN110240627A (en) * | 2019-07-19 | 2019-09-17 | 浙江长生鸟健康科技股份有限公司 | A method of small active peptides being extracted from pearl powder using complex enzyme |
-
2020
- 2020-01-16 CN CN202010049744.8A patent/CN111184744B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016112722A1 (en) * | 2015-01-15 | 2016-07-21 | 欧诗漫生物股份有限公司 | Preparation method for conchiolin, and water-soluble conchiolin and acid-soluble conchiolin prepared by using method |
| CN106538991A (en) * | 2016-11-01 | 2017-03-29 | 福建农林大学 | A kind of method that ultrasonic acidolysis pre-treat biological prepares kelp blood pressure product |
| CN110240627A (en) * | 2019-07-19 | 2019-09-17 | 浙江长生鸟健康科技股份有限公司 | A method of small active peptides being extracted from pearl powder using complex enzyme |
Non-Patent Citations (3)
| Title |
|---|
| detection and partial identification of protein in pearls formed in hyriopsis cumingii;Yang S C等;《African Journal of Biotechnology》;20111219;第10卷(第82期);19203-19219 * |
| 合浦珍珠提取物对金黄色葡萄球菌的体外抑菌活性及稳定性研究;刘鹏等;《食品工业科技》;20200731;第41卷(第14期);108-113 * |
| 合浦珠母贝源抗氧化肽的研究进展;吴燕燕等;《食品工业科技》;20171231;第38卷(第09期);339-343 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111184744A (en) | 2020-05-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4704377B2 (en) | Fermentation and culture method, plant fermented extract, plant fermented extract powder and blended plant fermented extract | |
| CN100512808C (en) | Externally applied compound amino acid prepn for promoting wound healing and its preparation process and application thereof | |
| EP2316464B1 (en) | Method of producing a dry earthworm powder | |
| TWI364284B (en) | Agonist for healing living organisms | |
| CN104163847B (en) | The preparation method of fly maggot active protein peptide and prepared fly maggot active protein peptide and application thereof | |
| CN110170047A (en) | A kind of antibacterial ointment of glusulase | |
| TWI825362B (en) | Use of lactobacillus fermentation product in preparation of external composition for enhancing skin wound healing | |
| CN111184744B (en) | Hepu pearl extract with antibacterial effect and preparation method thereof | |
| CN110590908B (en) | Micropterus-derived antibacterial peptide additive and preparation method thereof | |
| CN110272886A (en) | A kind of preparation method of snail enzyme solution and products thereof and application | |
| CN104840483A (en) | Medicine composition for treating skin wounds and medicinal preparation thereof | |
| US20220275419A1 (en) | Method for the preparation of low molecular weight porcine lympho-reticular polypeptides and formulations thereof | |
| DK157035B (en) | BIOLOGICALLY PURE MICROORGANISM STOCK AND PROCEDURE FOR THE PREPARATION OF A CULTIVATION PRODUCT THEREOF USED AS A SKIN PREPARATION | |
| CN104663585B (en) | Food toad return of spring worm breeding method and obtained pharmaceutical raw material | |
| CN111939215A (en) | Composition for treating bromhidrosis | |
| CN109999086A (en) | Treat pig polyserositis and arthritic Chinese medicine composition and preparation method thereof | |
| KR20160057183A (en) | Cosmetic composition containing the fermented extract of allium monanthum, allium scorodorpasum, allium tuberosum, allium fistulosum and scilla scilloides | |
| CN105801666B (en) | Preparation method and application of miichthys miiuy swim bladder protein peptide | |
| KR20040061707A (en) | The preparation method of the young antlers of the deer, deer blood or deer horns and the functional food and medicines using thereof | |
| CN102824369A (en) | Snake gall capsules and preparation method thereof | |
| CN113995775B (en) | Probiotics foot mask with foot protection effect and preparation method thereof | |
| CN106214704A (en) | A kind of Antheraea pernyi Geurin Meneville extract, large-scale method for producing and application | |
| CN107156746B (en) | Coconut honey and preparation method thereof | |
| CN116808115A (en) | Application of ampelopsis grossedentata extract in preparation of antibacterial and/or repairing products | |
| CN116115701A (en) | Antibacterial and virus-killing combined liquid and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |