CN103766343A - Tri-poly-glucosamine to hepta-poly poly-glucosamine monomers used as chitinase inhibitors - Google Patents
Tri-poly-glucosamine to hepta-poly poly-glucosamine monomers used as chitinase inhibitors Download PDFInfo
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- CN103766343A CN103766343A CN201410013553.0A CN201410013553A CN103766343A CN 103766343 A CN103766343 A CN 103766343A CN 201410013553 A CN201410013553 A CN 201410013553A CN 103766343 A CN103766343 A CN 103766343A
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- chitinase
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Abstract
The invention provides an oligomeric glucosamine monomer used as a chitinase inhibitor, belongs to the technical field of enzyme inhibitors, and particularly relates to an application of taking the oligomeric glucosamine monomer as the chitinase inhibitor. The oligomeric glucosamine is a tri-poly-glucosamine monomer, a tetra-poly-glucosamine monomer, a penta-poly-glucosamine monomer, a hexa-poly-glucosamine monomer and a hepta-poly poly-glucosamine monomer; and an oligomeric glucosamine water solution can be used as the chitinase inhibitor. The invention provides the natural product oligomeric glucosamine monomer which is used as the chitinase inhibitor; and the oligomeric glucosamine monomer has good water solubility, is safe and non-toxic and has a wide application prospect.
Description
Technical field
The invention belongs to enzyme inhibitor technical field, relate to a kind of 3 poly-, 4 poly-, 5 poly-, 6 poly-and 7 Chitosans as chitinase inhibitors.
Background technology
Chitin is the linear polysaccharide that N-Acetyl-D-glucosamine is formed by connecting with β-Isosorbide-5-Nitrae-glycosidic bond, is that natural world content is only second to cellulosic natural high polymer.Chitinase (EC3.2.1.14) is the key enzyme of chitin hydrolytic process, is responsible for poly-height chitin to be hydrolyzed to chitin oligo saccharide.Chitinase belongs to glycosyl hydrolase 18 families and 19 families, is distributed widely in microorganism, plant and animal body, is responsible for different physiological functions.Bacterium and part fungi utilize chitinase hydrolysis chitin as Carbon and nitrogen sources; Arthropods and part fungi utilize chitinase hydrolysis self to contain chitinous structure to meet the needs that grow; The defence of plant utilization chitinase is containing the infringement of chitin biology; Protozoa and part Mycophyta pathogene utilize chitinase to assist the invasion to host; Mammal chitinase participates in the process such as elementary immunity and inflammatory reaction to pathogene.Therefore, exploitation micromolecular inhibitor is used for the activity of chitinase and is with a wide range of applications.
Oligomerization gucosamine is to be formed by connecting by β-Isosorbide-5-Nitrae-glycosidic bond by gucosamine, has the advantage such as good water solubility, safety non-toxic.Oligomerization gucosamine, because of the particularity of its structure, has a wide range of applications at aspects such as nutrition and health care, livestock-raising, plant growth regulating and disease resistances.The oligomerization gucosamine that is part deacetylation of application at present, up to the present, the oligomerization gucosamine monomer of deacetylation does not have report as the application of chitinase inhibitors completely.
Summary of the invention
The invention provides the application as chitinase inhibitors of 3 poly-, 4 poly-, 5 poly-, 6 poly-and 7 Chitosans.
The scheme that technical solution problem of the present invention adopts is as follows:
As 3 poly-, 4 poly-, 5 poly-, 6 poly-and 7 Chitosans of chitinase inhibitors, called after (GlcN)
3to (GlcN)
7.
The invention provides 3 poly-, 4 poly-, 5 poly-, the 6 poly-and 7 polydextrose amine aqueous solutions external activity as chitinase inhibitors.3 poly-, 4 poly-, 5 poly-, 6 poly-and 7 polydextrose amine aqueous solutions all can suppress the activity of chitinase in vitro.
That the present invention also provides is 3 poly-, 4 poly-, 5 poly-, 6 poly-and 7 polydextrose amine aqueous solutions are as the activity in vivo of chitinase inhibitors.3 poly-, 4 poly-, 5 poly-, 6 poly-and 7 polydextrose amine aqueous solutions all can suppress the activity of chitinase in vivo, thereby cause insect normally not cast off a skin and dead.
Embodiment
Describe specific embodiments of the invention in detail below in conjunction with technical scheme.
The activity that embodiment 13 is poly-, 4 poly-, 5 poly-, 6 poly-and 7 Chitosans suppress chitinase in vitro
The chitinase of selecting is Asiatic corn borer chitinase OfChtI, mankind's chitinase HsCht and serratia marcescens chitinase SmChiA and SmChiB.Enzyme activity determination is selected p-nitrophenol-N, and N '-di-N-acetylchitobiose (pNP-β-(GlcNAc)
2) be substrate, there is characteristic absorption owing to generating free p-nitrophenol (pNP) after chitinase hydrolysis sugar glycosidic bond at 405nm, adopt 96 orifice plates to utilize microplate reader to measure enzyme activity.In separation and purification process, the mensuration of vigor is selected to 60 μ L reaction systems, in 96 orifice plates, mix a certain amount of chitinase and survey enzyme buffer solution (20mM NaH alive
2pO
4, pH6.0) totally 58.8 μ L, and 1.2 μ L5mM pNP-β-(GlcNAc)
2, after room temperature incubation certain hour, add 60 μ L0.5M Na
2cO
3cessation reaction, measures absorption value in 405nm.3 poly-, 4 poly-, 5 poly-, 6 inhibition activity poly-and 7 Chitosans obtain utilizing above-mentioned measuring method for activity to measure after 0～1000 μ M concentration gradient by dilution in 96 orifice plates.
Experimental result is as shown in table 1.
Table 13 is poly-, 4 poly-, 5 poly-, the 6 poly-and ICs of 7 Chitosans to chitinase
50value
| The μ M of unit | OfChtI | HsCht | SmChiA | SmChiB |
| (GlcN) 3 | 776.0 | 1610.0 | 1700.2 | 560.3 |
| (GlcN) 4 | 267.8 | 637.4 | 586.5 | 298.8 |
| (GlcN) 5 | 16.5 | 163.1 | 385.0 | 116.4 |
| (GlcN) 6 | 14.5 | 69.5 | 311.6 | 72.8 |
| (GlcN) 7 | 11.4 | 37.8 | 306.1 | 65.7 |
The activity that embodiment 23 is poly-, 4 poly-, 5 poly-, 6 poly-and 7 Chitosans suppress chitinase in vivo
Select Asiatic corn borer 5 instar larvaes as experiment material in body, each 45 of experimental group and control group.Every larva is injected 5 μ L with micro syringe at middle preabdomen.Experimental group injection concentration is 3 poly-, 4 poly-, 5 poly-, the 6 poly-and 7 polydextrose amine aqueous solutions of 10mg/ml, control group injection deionized water.Corn borer after treatment is fed with artificial feed, after pupating, observes lethality.
Experimental result is as shown in table 2, and dead phenotype is in the process of pupating and can not normally casts off a skin.
Table 23 is poly-, 4 poly-, 5 poly-, the 6 poly-and activity in vivo of 7 Chitosans to chitinase
| ? | (GlcN) 3 | (GlcN) 4 | (GlcN) 5 | (GlcN) 6 | (GlcN) 7 |
| Lethality (%) | 20% | 25% | 75% | 90% | 90% |
Claims (3)
1. the 3-7 polydextrose amine monomers as chitinase inhibitors, it is characterized in that, described oligomerization gucosamine monomer is β-1,3 poly-, 4 poly-, 5 poly-, 6 poly-or 7 Chitosans that 4-glycosidic bond connects, using above-mentioned oligomerization gucosamine monomer solution as chitinase inhibitors.
2. a kind of 3 poly-, 4 poly-, 5 poly-, 6 poly-and 7 Chitosans as chitinase inhibitors according to claim 1, is characterized in that, described chitinase is the chitinase of insect, vertebrate and bacterial origin.
3. a kind of 3 poly-, 4 poly-, 5 poly-, 6 poly-and 7 Chitosans as chitinase inhibitors according to claim 1 and 2, is characterized in that, described chitinase inhibitors is as insecticide, bactericide and treatment human diseases medicine.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107223663A (en) * | 2017-04-24 | 2017-10-03 | 大连理工大学 | Kasugarnycin and its application that derivative is chitinase inhibitors |
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| US6492350B2 (en) * | 2000-01-27 | 2002-12-10 | Jdc (Hawaii) Inc. | Chitin oligosaccharides and/or chitosan oligosaccharides for preventing or treating common cold or treating pain |
| CN1680569A (en) * | 2005-01-11 | 2005-10-12 | 苏理 | Production of chitose from fusarium solani |
| CN101619082A (en) * | 2008-07-01 | 2010-01-06 | 中国科学院大连化学物理研究所 | Method for separating and purifying chitosan oligosaccharide monomer |
| CN102174064A (en) * | 2011-03-22 | 2011-09-07 | 连云港海康生物科技有限公司 | Method for preparing chitosan oligosaccharide monomers by gel chromatography |
| CN102302037A (en) * | 2010-05-09 | 2012-01-04 | 海南正业中农高科股份有限公司 | Bactericide pesticide composition containing chitosan oligosaccharide |
| CN103081911A (en) * | 2013-01-10 | 2013-05-08 | 大连理工大学 | Application of chitosan oligosaccharide as insect chitinase inhibitor and pesticide |
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2014
- 2014-01-10 CN CN201410013553.0A patent/CN103766343A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6492350B2 (en) * | 2000-01-27 | 2002-12-10 | Jdc (Hawaii) Inc. | Chitin oligosaccharides and/or chitosan oligosaccharides for preventing or treating common cold or treating pain |
| CN1680569A (en) * | 2005-01-11 | 2005-10-12 | 苏理 | Production of chitose from fusarium solani |
| CN101619082A (en) * | 2008-07-01 | 2010-01-06 | 中国科学院大连化学物理研究所 | Method for separating and purifying chitosan oligosaccharide monomer |
| CN102302037A (en) * | 2010-05-09 | 2012-01-04 | 海南正业中农高科股份有限公司 | Bactericide pesticide composition containing chitosan oligosaccharide |
| CN102174064A (en) * | 2011-03-22 | 2011-09-07 | 连云港海康生物科技有限公司 | Method for preparing chitosan oligosaccharide monomers by gel chromatography |
| CN103081911A (en) * | 2013-01-10 | 2013-05-08 | 大连理工大学 | Application of chitosan oligosaccharide as insect chitinase inhibitor and pesticide |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107223663A (en) * | 2017-04-24 | 2017-10-03 | 大连理工大学 | Kasugarnycin and its application that derivative is chitinase inhibitors |
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Application publication date: 20140507 |