CN111705010B - Application of halomonas in synthesizing biodegradable mulching film by utilizing straw and kitchen waste grease - Google Patents
Application of halomonas in synthesizing biodegradable mulching film by utilizing straw and kitchen waste grease Download PDFInfo
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- CN111705010B CN111705010B CN202010103384.5A CN202010103384A CN111705010B CN 111705010 B CN111705010 B CN 111705010B CN 202010103384 A CN202010103384 A CN 202010103384A CN 111705010 B CN111705010 B CN 111705010B
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- 239000010902 straw Substances 0.000 title claims abstract description 68
- 239000010806 kitchen waste Substances 0.000 title claims abstract description 35
- 239000004519 grease Substances 0.000 title claims abstract description 28
- 241000206596 Halomonas Species 0.000 title claims abstract description 11
- 230000002194 synthesizing effect Effects 0.000 title claims abstract description 9
- 229920000728 polyester Polymers 0.000 claims abstract description 41
- 239000000126 substance Substances 0.000 claims abstract description 31
- 239000002362 mulch Substances 0.000 claims abstract description 29
- 241001653918 Halomonas sp. Species 0.000 claims abstract description 23
- 239000001963 growth medium Substances 0.000 claims abstract description 20
- 239000006228 supernatant Substances 0.000 claims abstract description 18
- 239000002689 soil Substances 0.000 claims abstract description 16
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims abstract description 12
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 claims abstract description 12
- 238000000034 method Methods 0.000 claims abstract description 12
- 239000002245 particle Substances 0.000 claims abstract description 10
- 239000003513 alkali Substances 0.000 claims abstract description 6
- 229910000019 calcium carbonate Inorganic materials 0.000 claims abstract description 6
- 239000004408 titanium dioxide Substances 0.000 claims abstract description 6
- AXKZIDYFAMKWSA-UHFFFAOYSA-N 1,6-dioxacyclododecane-7,12-dione Chemical compound O=C1CCCCC(=O)OCCCCO1 AXKZIDYFAMKWSA-UHFFFAOYSA-N 0.000 claims abstract description 3
- KKEYFWRCBNTPAC-UHFFFAOYSA-L terephthalate(2-) Chemical compound [O-]C(=O)C1=CC=C(C([O-])=O)C=C1 KKEYFWRCBNTPAC-UHFFFAOYSA-L 0.000 claims abstract description 3
- 239000000463 material Substances 0.000 claims description 18
- 239000000203 mixture Substances 0.000 claims description 14
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 12
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 12
- 239000000843 powder Substances 0.000 claims description 11
- 238000005406 washing Methods 0.000 claims description 11
- 238000000855 fermentation Methods 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 239000013585 weight reducing agent Substances 0.000 claims description 9
- 230000004151 fermentation Effects 0.000 claims description 8
- 238000004321 preservation Methods 0.000 claims description 8
- 230000015556 catabolic process Effects 0.000 claims description 7
- 238000012258 culturing Methods 0.000 claims description 7
- 238000006731 degradation reaction Methods 0.000 claims description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 6
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 229920005586 poly(adipic acid) Polymers 0.000 claims description 5
- 241001052560 Thallis Species 0.000 claims description 4
- 229940041514 candida albicans extract Drugs 0.000 claims description 4
- 238000005119 centrifugation Methods 0.000 claims description 4
- 238000010096 film blowing Methods 0.000 claims description 4
- 238000002329 infrared spectrum Methods 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 229920001707 polybutylene terephthalate Polymers 0.000 claims description 4
- BHDWNGYXLQBZKS-UHFFFAOYSA-N sodium;chloroform;hypochlorite Chemical compound [Na+].Cl[O-].ClC(Cl)Cl BHDWNGYXLQBZKS-UHFFFAOYSA-N 0.000 claims description 4
- 230000001954 sterilising effect Effects 0.000 claims description 4
- 239000012138 yeast extract Substances 0.000 claims description 4
- WSQZNZLOZXSBHA-UHFFFAOYSA-N 3,8-dioxabicyclo[8.2.2]tetradeca-1(12),10,13-triene-2,9-dione Chemical compound O=C1OCCCCOC(=O)C2=CC=C1C=C2 WSQZNZLOZXSBHA-UHFFFAOYSA-N 0.000 claims description 3
- 239000005708 Sodium hypochlorite Substances 0.000 claims description 3
- 239000001569 carbon dioxide Substances 0.000 claims description 3
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 3
- 238000000354 decomposition reaction Methods 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 239000003864 humus Substances 0.000 claims description 3
- VUZPPFZMUPKLLV-UHFFFAOYSA-N methane;hydrate Chemical compound C.O VUZPPFZMUPKLLV-UHFFFAOYSA-N 0.000 claims description 3
- 238000007873 sieving Methods 0.000 claims description 3
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims description 3
- 230000007480 spreading Effects 0.000 claims description 3
- 238000003892 spreading Methods 0.000 claims description 3
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 claims description 2
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 claims description 2
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 claims description 2
- 239000005642 Oleic acid Substances 0.000 claims description 2
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 claims description 2
- 239000010775 animal oil Substances 0.000 claims description 2
- 238000005286 illumination Methods 0.000 claims description 2
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 claims description 2
- 238000009629 microbiological culture Methods 0.000 claims description 2
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 claims description 2
- 229920000642 polymer Polymers 0.000 claims description 2
- 238000001179 sorption measurement Methods 0.000 claims description 2
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 claims description 2
- 235000015112 vegetable and seed oil Nutrition 0.000 claims description 2
- 239000008158 vegetable oil Substances 0.000 claims description 2
- 230000003834 intracellular effect Effects 0.000 claims 3
- 238000000071 blow moulding Methods 0.000 claims 2
- 230000001580 bacterial effect Effects 0.000 claims 1
- 239000008187 granular material Substances 0.000 claims 1
- 239000013067 intermediate product Substances 0.000 claims 1
- 238000002844 melting Methods 0.000 claims 1
- 230000008018 melting Effects 0.000 claims 1
- 108090000623 proteins and genes Proteins 0.000 claims 1
- 230000001105 regulatory effect Effects 0.000 claims 1
- 238000002791 soaking Methods 0.000 claims 1
- 238000003756 stirring Methods 0.000 claims 1
- 239000003921 oil Substances 0.000 description 11
- 235000019198 oils Nutrition 0.000 description 11
- 229910052799 carbon Inorganic materials 0.000 description 9
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 7
- 239000002699 waste material Substances 0.000 description 7
- 241000894006 Bacteria Species 0.000 description 6
- 230000008901 benefit Effects 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- 229920006238 degradable plastic Polymers 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 238000001125 extrusion Methods 0.000 description 3
- 238000005469 granulation Methods 0.000 description 3
- 230000003179 granulation Effects 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 239000005014 poly(hydroxyalkanoate) Substances 0.000 description 3
- 229920000903 polyhydroxyalkanoate Polymers 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 150000001721 carbon Chemical group 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000012844 infrared spectroscopy analysis Methods 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 239000003208 petroleum Substances 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 239000008399 tap water Substances 0.000 description 2
- 235000020679 tap water Nutrition 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- LQLQDKBJAIILIQ-UHFFFAOYSA-N Dibutyl terephthalate Chemical compound CCCCOC(=O)C1=CC=C(C(=O)OCCCC)C=C1 LQLQDKBJAIILIQ-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 241000206595 Halomonas elongata Species 0.000 description 1
- 238000004566 IR spectroscopy Methods 0.000 description 1
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 239000002154 agricultural waste Substances 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000002856 computational phylogenetic analysis Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000004626 scanning electron microscopy Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000010802 sludge Substances 0.000 description 1
- 238000004158 soil respiration Methods 0.000 description 1
- 238000010183 spectrum analysis Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000002834 transmittance Methods 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J5/00—Manufacture of articles or shaped materials containing macromolecular substances
- C08J5/18—Manufacture of films or sheets
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/62—Carboxylic acid esters
- C12P7/625—Polyesters of hydroxy carboxylic acids
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2300/00—Characterised by the use of unspecified polymers
- C08J2300/16—Biodegradable polymers
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2367/00—Characterised by the use of polyesters obtained by reactions forming a carboxylic ester link in the main chain; Derivatives of such polymers
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2467/00—Characterised by the use of polyesters obtained by reactions forming a carboxylic ester link in the main chain; Derivatives of such polymers
- C08J2467/02—Polyesters derived from dicarboxylic acids and dihydroxy compounds
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K3/00—Use of inorganic substances as compounding ingredients
- C08K3/18—Oxygen-containing compounds, e.g. metal carbonyls
- C08K3/20—Oxides; Hydroxides
- C08K3/22—Oxides; Hydroxides of metals
- C08K2003/2237—Oxides; Hydroxides of metals of titanium
- C08K2003/2241—Titanium dioxide
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K3/00—Use of inorganic substances as compounding ingredients
- C08K3/18—Oxygen-containing compounds, e.g. metal carbonyls
- C08K3/24—Acids; Salts thereof
- C08K3/26—Carbonates; Bicarbonates
- C08K2003/265—Calcium, strontium or barium carbonate
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- General Chemical & Material Sciences (AREA)
- Materials Engineering (AREA)
- Polymers & Plastics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Separation, Recovery Or Treatment Of Waste Materials Containing Plastics (AREA)
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Abstract
The invention discloses a method for synthesizing a biodegradable mulch film by Halomonas sp.ZY-1 utilizing straw and kitchen waste grease. And (3) treating the straw by alkali liquor to obtain straw supernatant, and preparing an MS culture medium by the straw supernatant and kitchen waste grease in a volume ratio of 35:1-40:1. The halomonas is fermented by an MS culture medium to obtain polyester substances, 60-80 parts by mass of the polyester substances, 30-50 parts by mass of poly (butylene adipate)/terephthalate), 0.5-1 part by mass of calcium carbonate and 0.2-0.5 part by mass of titanium dioxide are uniformly mixed, melted and extruded into particles, and the particles are blown into films, wherein the tensile strength of the films is 16-18MPa, and the films have biodegradability and toughness and can be used for producing degradable mulching films. The weight of the mulching film is reduced by about 50% when the mulching film is buried in soil with the depth of 20-25cm at 20-25 ℃ and reduced by about 0.5% when the mulching film is buried in the air for 35 d.
Description
Technical Field
The invention belongs to the field of agricultural environmental protection, and particularly relates to a method for synthesizing a biodegradable mulch film by utilizing Halomonas sp.zy-1 and straw and kitchen waste grease.
Background
Drought and low temperature are one of the main obstacles for sustainable development of northern agriculture in China, and seriously affect the development of agricultural economy in northeast areas. The laying of the mulch film makes great contribution to solving the problems of heat preservation and drought resistance of crop growth, but the mulch film made of traditional petrochemical plastics has stable chemical structure and is difficult to degrade in a short time, influences soil respiration, pollutes cultivated land and leads to crop yield reduction. At present, the degradable plastic produced by microorganisms mainly uses exquisite carbon sources such as glucose, rhamnose, molasses and the like as fermentation substrates, and can use less crude carbon sources such as straw, kitchen waste grease and the like. When the microbes are used for producing the degradable plastics, the temperature strain is used, high-temperature high-pressure sterilization is needed, and the extremely microbes are rarely used for producing the degradable plastics. The advantage of Halomonas sp.zy-1 is mainly expressed in that the straw supernatant and the treated kitchen waste grease can be used as carbon to produce degradable plastic PHA under the condition of high-salt and high-alkali environment (60 g/L NaCl, pH 9). Other strains for PHA production use mainly waste water, waste oil, activated sludge, but rarely straw carbon sources, so the strain has great advantages in substrate selection and waste utilization. And the weight reduction rate of the degradable biological mulch film in the dry air is 0.5%, the weight reduction rate of 35d reaches 50% in the soil which is deeply buried by 20-30cm, the degradation rate can be accelerated along with the increase of illumination, the soil humus is increased by the biodegradable mulch film, the soil hardening is effectively relieved, and the degradable biological mulch film is finally decomposed into carbon dioxide and water.
Disclosure of Invention
The invention aims to solve the problem that the existing biodegradable mulching film cannot utilize microorganisms, straws and kitchen waste grease. The method for producing the biodegradable mulching film by using the straw and the kitchen waste grease to synthesize the ZY-1 by using the Halomonas sp further optimizes the material chemical property of the biodegradable mulching film to prepare the biodegradable mulching film with the elasticity and the transmittance meeting the conditions. The method for synthesizing the biodegradable mulch film is green, pollution-free, low in cost and simple in operation.
The invention relates to a Halomonas which is Halomonas (Halomonas sp.) ZY-1 and is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms, wherein the preservation address is No.1, no. 3 of North Chen West Lu in the Korean area of Beijing, the preservation date is 2018, 11 and 22 days, and the preservation number is CGMCC No.16773.
The application of the halomonas strain disclosed by the invention is used for preparing biodegradable mulching films.
The preparation method of the biodegradable mulching film comprises the following steps:
(1) Washing the straw with tap water until no soil exists, drying at 50-60 ℃, crushing by a crusher, and sieving by a 100-200 mesh sieve to obtain straw powder. Placing 50-75g of straw powder into 1L of 2mol/L NaOH solution, treating at 105-115 ℃ for 1-2h, centrifuging at 4000-6000r/min for 10min after cooling, and collecting straw supernatant.
(2) Preparing MS culture medium comprising straw supernatant 750-800mL/L, kitchen waste oil 17-20mL/L, yeast extract 5g/L, na 2 HPO 4 ·12H 2 O 10-15g/L、KH 2 PO 4 1-2g/L、NH 4 Cl 1-2g/L、MgSO 4 0.1-1.5 g/L、Fe(III)-NH 4 -Citrate 0.05-0.1g/L、CaCl 2 ·2H 2 O 0.02-0.1g/L、ZnSO 4 ·7H 2 O0.1-0.5 g/L, naCI g/L, and pH is adjusted to 8.0-10.
(3) Inoculating 1-5% Halomonas sp.ZY-1 strain into LB culture medium (pH 8.0-10, naCI 60 g/L), culturing at 36-38deg.C with shaking incubator at 140-160r/min for 12-14h, inoculating 1-5% seed solution into MS culture medium, and culturing at 36-38deg.C with shaking incubator at 140-160r/min for 70-90h.
(4) Centrifuging the fermentation broth at 4000-6000r/min for 10min to collect thalli, pre-cooling at-20deg.C for 12-14h, and lyophilizing in a lyophilizing machine for 12-24h. Sequentially washing with distilled water, centrifuging, washing with acetone, centrifuging, washing with water again, centrifuging at 4000-6000r/min for 10min, placing thallus into a conical flask, and adding chloroform: centrifuging 4000-6000r/min for 10min after uniformly mixing the mixed solution with 3:1 sodium hypochlorite, sucking the lower chloroform phase, spreading in a clean container, adding pre-cooled 10-15ml of 95% ethanol, and separating out polyester substances.
(5) 60-80 parts of polyester substances, 30-50 parts of poly (adipic acid)/butylene terephthalate, 0.5-1 part of calcium carbonate and 0.2-0.5 part of titanium dioxide are put into a high-speed mixer and stirred for 15-20min at the rotating speed of 800-1000r/min, and the mixture is uniformly mixed.
(6) The obtained mixture is sent into a double-screw extruder, the screw rotating speed is 160r/min, the feeding speed is 20r/min, extrusion granulation is carried out at 160-180 ℃ to obtain mixture particles, and the mixture particles are blown to form a film on a film blowing unit to prepare the biodegradable mulch film.
The Halomonas sp.zy-1 used in the invention is obtained by screening and separating a sample from saline-alkali soil in a large area of Daqing city, the Genebank accession number is MH428215, and the similarity of the strain and Halomonas elongata is 98% through phylogenetic tree analysis and identification, and the strain is named Halomonas sp.zy-1, as shown in figure 1. The microbial strain is preserved in China center for type culture collection (CGMCC) in 11 and 22 days of 2018, and the address is in 1 # 3 of North Silu of the Chaoyang area of Beijing city, and the preservation number is CGMCC No.16773.
The invention has the advantages that:
(1) Strain selection advantages: the halophilic bacteria (Halomonas sp.) ZY-1 is halophilic bacteria, survives under the environmental conditions of high salt and high alkali (pH 8.0-10, 60g/L NaCl), but the non-halophilic bacteria cannot survive in the special environment, and the characteristics that halophilic bacteria can inhibit the growth of mixed bacteria by utilizing the growth of halophilic bacteria in the high salt and high alkali environment are utilized, so that the investment of high-energy sterilization and fermentation tanks in production is reduced, the investment of high-pressure steam sterilization energy sources and closed fermentation equipment is saved, and the cost investment in the production process is effectively reduced.
(2) The raw material selection has the advantages: the waste agricultural waste straw and kitchen waste grease are used for replacing an exquisite carbon source, so that the cost of raw materials for production is reduced, and resources are saved; the biodegradable mulching film synthesized by waste resources replaces petroleum-based plastics, so that white pollution can be avoided, the environment is protected, the conversion from petrochemical economy to green recycling economy is accelerated, and the low-cost, environment-friendly and degradable material is synthesized.
(3) The biodegradable mulching film has the advantage of no pollution after decomposition: the weight reduction rate of the synthesized biodegradable mulching film is about 50% when the mulching film is deeply buried in soil with 20-30cm of cultivated land at 20-25 ℃; the weight reduction rate of the 35d film is 0.1% under the condition of dry room temperature, and the product is suitable for popularization and application on agricultural mulching films, and lays a foundation for the development of green recyclable agriculture economy.
In conclusion, the biodegradable mulching film synthesized by straw and kitchen waste oil replaces the petroleum-based mulching film, so that the white pollution can be avoided, the environment is protected, and the agricultural farmland is protected
Drawings
FIG. 1 is a phylogenetic tree of Halomonas sp.ZY-1;
FIG. 2 is a natural degradation of a biodegradable mulch;
FIG. 3 is a graph showing the yield analysis of Halomonas sp.ZY-1 for synthesizing polyester based materials from straw;
FIG. 4 is an analysis of the output of Halomonas sp.ZY-1 for synthesizing polyester based materials from straw and kitchen waste oil;
FIG. 5 shows the transmission electron microscope observation of biomass and polyester based materials of Halomonas sp.ZY-1 utilizing straw and kitchen waste grease;
FIG. 6 shows the synthesis of polyester based materials from straw and kitchen waste grease by Halomonas sp.ZY-1;
FIG. 7 is a Scanning Electron Microscope (SEM) analysis of different polyesters;
FIG. 8 is an infrared spectroscopic analysis (FTIR) of different polyesters;
FIG. 9 is a graph showing the tensile strength of synthetic films of varying poly (adipic acid)/poly (butylene terephthalate) content;
FIG. 10 shows a synthetic bio-mulch by bio-fermentation and modification of straw and kitchen waste grease.
Detailed Description
The experimental methods in the following examples are conventional methods unless otherwise specified. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Example 1 Natural degradation of biodegradable mulch film
The research is to simulate the degradation condition of the biodegradable mulch film under natural conditions, and examine the degradation capacity of the biodegradable mulch film by taking the weight reduction rate of the biodegradable mulch film buried in soil as an index.
(1) Taking 3 parts of prepared biodegradable mulch film sample, wherein each part is 20-30mg, and marking as initial mass M 0 。
(2) And (3) respectively placing the biodegradable mulch film samples in cultivated lands with the depth of 15-20cm at the temperature of 20-25 ℃, taking out the samples every 7d to remove surface soil, placing the samples in an oven with the temperature of 40-55 ℃ for drying treatment, and weighing and marking the samples as Mn.
(3) The weight loss rate of the biodegradable mulch film is calculated as follows:
the degradation condition is as shown in figure 2, the weight reduction rate of 35d in dry air is 0.5%, the degradable mulch film is deeply buried in 15-20cm farmland soil at 20-25 ℃, the weight reduction rate of 35d reaches 40-50%, the humus of the soil can be increased after decomposition, the problem of soil hardening is relieved, and finally the degradable mulch film is converted into carbon dioxide and water.
Example 2 Synthesis of polyester-based Material from straw by Halomonas sp.ZY-1
(1) The crushed straws are sieved by a 100-200 mesh sieve, 50-75g of straw powder is soaked in 1L of 1-2mol/L NaOH solution, the high temperature and high pressure treatment is carried out for 1-2h, and the centrifugation is carried out for 10min at 4000-6000r/min to collect straw supernatant.
(2) The straw supernatant is used as a carbon source MS culture medium, the Halomonas sp.ZY-1 is activated by an LB culture medium overnight, the inoculation amount is 1-5%, the activated seed liquid is inoculated into the MS culture medium, and the culture is carried out for 60-96h at the temperature of 36-38 ℃ and the rotating speed of a shake incubator of 140-160 r/min.
(3) The chloroform sodium hypochlorite method is adopted to extract the polyester substances, and the yield change of fermentation in different periods is observed, as shown in figure 3, and the yield of the polyester substances reaches the maximum value of 0.0805g/L during 84h of fermentation.
Example 3 Synthesis of polyester-based Material from Rice straw and kitchen waste oil Using Halomonas sp.ZY-1
According to the analysis result of the yield of the polyester-based material in example 2, the yield of the polyester-based material synthesized from the straw was low, and the film forming property of the polyester-based material was poor. Adding kitchen waste oil (90-92% oleic acid, 3-4% triglyceride, 2-3% vegetable oil, and 2-3% animal oil) for improvement, and preparing MS culture medium with straw and kitchen waste oil as carbon.
(1) Preparing a gradient MS culture medium of straw and kitchen waste grease: 800mL/L straw supernatant, yeast extract 5g/L, na 2 HPO 4 12H 2 O 10-15g/L、KH 2 PO 4 1-2g/L、NH 4 Cl 1-2g/L、MgSO 4 0.1-1.5g/L、 Fe(III)-NH 4 -Citrate 0.05-0.1g/L、CaCl 2 ·2H 2 O 0.02-0.1g/L、ZnSO 4 ·7H 2 O0.1-0.5 g/L, and kitchen waste grease is 5mL/L, 10mL/L, 15mL/L, 20mL/L, 25mL/L and 30mL/L respectively.
(2) Inoculating Halomonas sp.ZY-1 seed solution activated overnight in LB culture medium into MS culture medium at 1-5% inoculum size, and culturing at 36-38deg.C in shake incubator at 140-160r/min for 80-86 hr.
(3) The change of the yield of the kitchen waste grease with different contents is analyzed by adopting a chloroform sodium hypochlorite method to extract the polyester substances, as shown in figure 4, under the condition that the volume of the straw supernatant fluid is fixed, the yield of the polyester substances is gradually increased along with the increase of the content of the kitchen waste grease, and when the concentration of the kitchen waste grease is 20mL/L, namely, the ratio of the straw supernatant fluid to the kitchen waste grease is 40:1, the highest yield of the polyester substances is 1.57g/L.
As can be seen by a transmission electron microscope in FIG. 5, white particles in Halomonas sp.ZY-1 cells are polyester-based substances, and the extracted polyester-based substances are in a semitransparent film shape as shown in FIG. 6. Compared with the straw synthesized product, the toughness is improved, the film forming capability is improved, and the transparency is improved.
Example 4 Scanning Electron Microscopy (SEM) and Infrared Spectroscopy (FTIR)
(1) And taking a proper amount of straw to synthesize polyester substances and taking the straw and kitchen waste grease to synthesize the polyester substances, and observing the surface morphology of the material by a scanning electron microscope.
(2) A certain amount of PHB standard (Sigma) is weighed respectively,Synthesizing polyester substances from straw and waste oil from kitchen, tabletting with potassium bromide, and testing at 4000-500cm -1 Infrared spectroscopic analysis was performed.
The surface morphology of the material observed by the scanning electron microscope is as shown in figure 7, compared with the surface morphology of the material observed by the scanning electron microscope under 500 times of electron microscope, the surface of the polyester substance synthesized by the straw is rough, and the surface of the polyester substance synthesized by the waste oil of the straw kitchen is flat; under 10000 times of electron microscope, the polyester substance synthesized by the straw has obvious band-shaped cracks, and the material synthesized by the waste oil of the straw kitchen has less relative cracks. The energy spectrum analysis shows that: the carbon atom content of the polyester substance synthesized by the straw is 70-72%, and the carbon atom content of the polyester substance synthesized by the straw and the kitchen waste grease is 73-77%.
As shown in FIG. 8, the infrared spectrum analysis shows that the two polyester substances are at 1450, 2980 and 1720cm -1 Where corresponds to CH 2 The groups CH, c=o have adsorption bands, respectively. Corresponding to the infrared spectrum of PHB standard, the polymer is polyhydroxyalkanoate.
Example 5 optimization of the Material Properties of biodegradable mulch
(1) Converting straw and kitchen waste grease into polyester substances by utilizing Halomonas sp.ZY-1 to prepare a biodegradable mulching film;
(2) Setting the ratio of poly adipic acid/butyl terephthalate as 20, 30, 40, 50 and 60 with different mass portions, and other components as follows: 80 parts by mass of polyester substance, 0.5 part by mass of calcium carbonate and 0.2 part by mass of titanium dioxide are put into a high-speed mixer and stirred for 15-20min at a rotating speed of 800-1000r/min, and uniformly mixed;
(3) The obtained mixture is sent into a double-screw extruder, the screw rotating speed is 160r/min, the feeding speed is 20r/min, and extrusion granulation is carried out at 160-180 ℃ to obtain mixture particles.
(4) The obtained mixture particles were blow molded on a film blowing machine to form a film, see fig. 10, and the tensile strength of the film was measured according to GB 2568.
As a result of measuring the tensile force, as shown in FIG. 9, when 50 parts by mass of poly (butylene adipate/terephthalate) was used, the obtained biodegradable mulch film had the highest strength and the tensile strength was 16MPa.
In order to achieve the above purpose, the technical scheme of the invention is as follows:
(1) Washing the straw with tap water until no soil exists, drying at 50-60 ℃, crushing by a crusher, and sieving by a 100-200 mesh sieve to obtain straw powder. Placing 50-75g of straw powder into 1L of 2mol/L NaOH solution, treating at 105-115 ℃ for 1-2h, centrifuging at 4000-6000r/min for 10min after cooling, and collecting straw supernatant.
(2) Preparing MS culture medium comprising straw supernatant 750-800mL/L, kitchen waste oil 17-20mL/L, yeast extract 5g/L, na 2 HPO 4 ·12H 2 O 10-15g/L、KH 2 PO 4 1-2g/L、NH 4 Cl 1-2g/L、MgSO 4 0.1-1.5 g/L、Fe(III)-NH 4 -Citrate 0.05-0.1g/L、CaCl 2 ·2H 2 O 0.02-0.1g/L、ZnSO 4 ·7H 2 O0.1-0.5 g/L, naCI g/L, and pH is adjusted to 8.0-10.
(3) Inoculating 1-5% Halomonas sp.ZY-1 strain into LB culture medium (pH 8.0-10, naCI 60 g/L), culturing at 36-38deg.C with shaking incubator at 140-160r/min for 12-14h, inoculating 1-5% seed solution into MS culture medium, and culturing at 36-38deg.C with shaking incubator at 140-160r/min for 70-90h.
(4) Centrifuging the fermentation broth at 4000-6000r/min for 10min to collect thalli, pre-cooling at-20deg.C for 12-14h, and lyophilizing in a lyophilizing machine for 12-24h. Sequentially washing with distilled water, centrifuging, washing with acetone, centrifuging, washing with water again, centrifuging at 4000-6000r/min for 10min, placing thallus into a conical flask, and adding chloroform: centrifuging 4000-6000r/min for 10min after uniformly mixing the mixed solution with 3:1 sodium hypochlorite, sucking the lower chloroform phase, spreading in a clean container, adding pre-cooled 10-15ml of 95% ethanol, and separating out polyester substances.
(5) 60-80 parts of polyester substances, 30-50 parts of poly (adipic acid)/butylene terephthalate, 0.5-1 part of calcium carbonate and 0.2-0.5 part of titanium dioxide are put into a high-speed mixer and stirred for 15-20min at the rotating speed of 800-1000r/min, and the mixture is uniformly mixed.
(6) The obtained mixture is sent into a double-screw extruder, the screw rotating speed is 160r/min, the feeding speed is 20r/min, extrusion granulation is carried out at 160-180 ℃ to obtain mixture particles, and the mixture particles are blown to form a film on a film blowing unit to prepare the biodegradable mulch film.
Claims (5)
1. The method for preparing the biodegradable mulching film by utilizing straw and kitchen waste grease by utilizing Halomonas sp.ZY-1 comprises the following steps:
step 1: crushing straw into straw powder, treating the straw powder by alkali liquor to obtain straw supernatant, and adding kitchen waste grease to ensure that the volume ratio of the straw supernatant to the kitchen waste grease is 35-40: 1, preparing an MS culture medium;
step 2: inoculating Halomonas sp ZY-1 into an MS culture medium prepared from straw supernatant and kitchen waste grease, fermenting and culturing for 70-90h to obtain bacterial cells, and breaking the wall by using a chloroform sodium hypochlorite method to extract intracellular polyester substances to obtain the biodegradable mulch;
step 3: modifying intracellular polyester substances, uniformly mixing polyester substances with poly (adipic acid)/butylene terephthalate, calcium carbonate and titanium dioxide, melting, extruding, and blow molding to obtain a film, thus obtaining the modified biodegradable mulching film;
the whole Halomonas is called Halomonas sp ZY-1, and is identified by the accession number MH428215 in Gene bank;
the straw supernatant is obtained by the following steps: crushing the straw by a crusher, sieving the crushed straw by a 100-200-mesh sieve to obtain straw powder, soaking 50-75g of straw powder in 1-2mol/L NaOH solution per 1L, treating the straw powder at high temperature and high pressure for 1-2h, centrifuging the straw powder at 4000-6000r/min for 10min, and collecting straw supernatant;
the kitchen waste grease comprises 90-92% of oleic acid, 3-4% of triglyceride, 2-3% of vegetable oil and 2-3% of animal oil;
the MS culture medium in the step 1 comprises 750-800mL/L of straw supernatant, 17-20mL/L of kitchen waste grease and 5g/L, na of yeast extract 2 HP0 4 12H 2 0 10～15g/L、KH 2 PO 4 1～2g/L、NH 4 Cl 1～2g/L、MgSO 4 0.1～1.5g/L、Fe(III)-NH 4 -Citrate 0.05～0.1g/L、CaCl 2 2H 2 0.02-0.1 g/L and ZnSO 4 ·7H 2 0.1-0.5 g/L; sterilizing the MS culture medium at 115 ℃ for 20min, and regulating the pH to 8-10;
the Halomonas (Halomonas sp.) ZY-1 in the step 1 is preserved in the China general microbiological culture Collection center, the preservation address is No.1, no. 3, and the preservation date is 22 days of 11 months in 2018, and the preservation number is CGMCC No.16773.
2. The method of claim 1, wherein the biodegradable mulch has a 35d weight reduction rate of 0.5% in dry air, is deeply buried in 20-30cm soil, has a 35d weight reduction rate of 50%, increases the surface temperature with the increase of illumination, increases the degradation rate, increases soil humus by intermediate products of decomposition of the biodegradable mulch, effectively relieves soil hardening, and finally decomposes into carbon dioxide and water.
3. The method according to claim 1, wherein in the step 2, the step of breaking the wall to extract the intracellular polyester material by using chloroform sodium hypochlorite method is characterized in that:
(1) Inoculating 1-5% Halomonas sp.ZY-1 seed solution into an MS culture medium, culturing for 70-90h at the temperature of 36-38 ℃ and the rotating speed of 140-160r/min, centrifuging the fermentation liquor at 4000-6000r/min for 10min, and collecting thalli;
(2) The thalli are placed in a freeze dryer for precooling for 12-14h at the temperature of minus 20 ℃, freeze-dried for 12-24h, and sequentially subjected to distilled water washing, centrifugation, acetone washing, centrifugation, water washing again and centrifugation, and centrifuged for 10min at 4000-6000 r/min;
(3) The cells were placed in a conical flask, and chloroform was added: sodium hypochlorite 3:1 (v/v), centrifuging for 10min at 4000-6000r/min after uniformly mixing, sucking the lower chloroform phase, spreading in a clean container, adding pre-cooled 10-15ml of 95% ethanol, and separating out polyester substances.
4. The method according to claim 1, wherein the obtained biodegradable mulch film in step 2 is found to be 14 by comparing the infrared spectrum with the PHB standard analysis50. 2980 and 1720cm -1 Where corresponds to CH 2 The CH and C=O groups are respectively provided with an adsorption band, which corresponds to the infrared spectrum of the PHB standard substance, and the polymer is PHB.
5. The method according to claim 1, wherein the specific step of synthesizing the modified biodegradable mulch film in the step 3 is to mix 60-80 parts by weight of polyester, 30-50 parts by weight of poly (butylene adipate)/terephthalate), 0.5-1 part by weight of calcium carbonate and 0.2-0.5 part by weight of titanium dioxide, stir the mixture in a high-speed mixer at a rotational speed of 800-1000r/min for 15-20min, send the mixture into a double-screw extruder at a screw rotational speed of 160r/min, extrude and granulate the mixture particles obtained at a feeding speed of 20r/min and a temperature of 160-180 ℃ to form a film by blow molding on a film blowing machine, and prepare the modified biodegradable mulch film.
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