41 results about "Nicotine degradation" patented technology

The invention discloses an Acinetobacter and application thereof. Production bacterial strain liquid is fermented and cultured to form corresponding preparation. The Production bacterial strain is the Acinetobacter (Acinetobacter sp.ND12); and the bacterial strain was preserved in China General Microbiological Culture Collection Center on November 5, 2009, and a preservation number is CGMCC No.3410. The bacterial strain has stronger nicotine degradation capacity and nicotine toxicity resistance capacity, and can be grown at the nicotine concentration of 3.5g / L; and by large-scale culturing of the bacterial strain, the pure nicotine can be degraded by using a whole cell of the bacterial strain as a biocatalyst, and meanwhile the content of the nicotine in tobacco is also reduced. The Acinetobacter and the application thereof have the advantages of capacities of adjusting the content of the nicotine in the tobacco raw material, improving availability of the tobacco, degrading the nicotine in smoke dust, reducing pollution of the nicotine to environment and the like.

The invention relates to a technology of biological degradation of nicotine in tobacco, in particular to a pseudomonad ZCJ bacterial strain applied to nicotine degradation of tobacco and a screening method and an application thereof. The pseudomonad ZCJ bacterial strain (Pseudomonas sp. ZCJ) was preserved in the China Center for Type Culture Collection on March, 30th, 2009 with a preservation number of CCTCC NO. M209064. The invention also discloses screening and application of the bacterial strain. In the invention, nicotine degradation bacterial strain screened from tobacco materials has relatively high biological activity and is applicable to degradation of nicotine in tobacco in a solid state, thus achieving the purpose of the degradation of the nicotine in tobacco.

The present invention discloses a quorum-sensing signal molecular preparation, which is prepared through the following steps: culturing alpha-proteobacterial, delta-proteobacterial and flavobacteria microorganisms respectively; taking culture solution or supernate made by centrifugalizing culture solution, and mixing with the volume proportion of 1-4:1:1-2; and cooling and drying. The quorum-sensing signal molecular preparation can precipitate nicotine degrading bacteria to colonize and constantly multiply in the contaminated environment, and can precipitate microbial populations in the environment to dynamically develop, so as to effectively improve nicotine removal and the comprehensive treatment efficiency of contaminants. The preparation can treat tobacco waste in cooperation with nicotine degrading bacteria and has the advantage of lasting action with one time of feeding. The preparation can greatly reduce the starting time for an active sludge reactor and accelerate the stabilization of the reactor. The operation is convenient, and both the cost and the maintenance expense can be reduced greatly. Therefore, the present invention provides an ideal treatment method for tobacco waste.

The invention relates to a method of biology degradation nicotine which uses (Ochrobactrum intermedium) DN2 as bacteria seed and uses the nicotine zymosis enzyme of the Ochrobactrum intermedium DN2 or Ochrobactrum intermedium DN2 to effect on the flue-cured tobacco leaf, pipe tobacco, industrial or agricultural tobacco leftover bits and pieces and tobacco residues and the extracting liquid of the above materials to do degradation to part of or all of the nicotine.

The invention provides a novel Shinella sp. HZN1 capable of effectively degrading nicotine and application thereof. The Shinella sp. HZN1 is preserved in China Center for Type Culture Collection; the address is 430072, Wuhan University, Wuhan, China; the preservation number is CCTCC No:M 2010154; and the preservation date is June 23rd, 2010. The nicotine-degrading bacterium can be directly thrown in a water body to degrade nicotine, and can safely, efficiently and quickly degrade the residual nicotine in the water body, soil and the like. Besides, the microbial inoculum containing the strain has the advantages of simple preparation process, low cost and convenient use, thereby having favorable application prospects.

The present invention relates to a novel tobacco leaf fermenting enzyme preparation production method which aims to solve the technical problems that how the quality of the tobacco leaf fermenting product is improved and how the reactivity protection of the tobacco leaf fermenting enzyme preparation is realized in the natural fermenting field of the tobacco leaf. The enzyme preparation consists of glucoseoxidase, chlorophyl oxidase, carotenoid oxidase, protease, and nicotine-degradation enzyme. Through the cell disruption of fresh leaves, (NH 4) 2 SO 4 is utilized to operate the second fractional precipitation to obtain crude enzyme fluid, an enzyme molecule adopts Ca 2 + and Mg 2 + to operate the metal ion exchange, to accomplish the molecule modification; a macro molecule combination modification is accomplished through adopting 0.01 percent of cane sugar low molecular polymer, thereby prolonging the half life period of an enzyme preparation, and obviously improving the high temperature resistant ability. The experimental result employed by the enzyme preparation indicates that the nicotine is decreased by 9.3 percent, the total nitrogen is decreased by 5.7 percent, the protein is decreased by 7.1 percent; cigarette smoke condensates are decreased by 8.4 percent, the tar content is decreased by 5.1 percent, cigarette smoke nicotine content is decreased by 28.0 percent, and the carbon is decreased monoxide by 1.6 percent. The enzyme preparation is employed when the tobacco is wet for the second time before defolat and redrying.

The invention provides a new nicotine-degrading bacteria-pseudomonas ZUTSKD and its application. The ZUTSKD (Pseudomonas sp.ZUTSKD) is stored in China Center for Type Culture Collection, the storing date was June 18th, 2007, the preservation no. is CCTCC NO:M 207083. The bacteria can degrade nicotine and generate such intermediate as 2, 3'-Bipyridine, cotinine, 3-(3, 4-dihydro-2H-pyrrol-5-y1)-Pyridine and 3-Pyridinecarboxylic acid, the max. nicotine concentration withstandable is 5.5 g / L. The invention can reduce the nicotine in waste cigarette residues, and is of certain application prospect in the industry.

The invention discloses a pseudomonas putida strain used for nicotine degradation with the preservation number being CGMCC NO.13055. The strain is obtained from a tobacco field soil sample and used for degrading nicotine. The invention further discloses an isolation and identification method of the pseudomonas putida strain used for nicotine degradation. The pseudomonas putida strain used for nicotine degradation has the beneficial effects that the strain has high biological activity, and is easy to culture and stable in genetic property, the nicotine degradation rate is high, the pseudomonas putida strain can be used for degrading the nicotine in tobacco or tobacco waste, and the nicotine in tobacco leaves is reduced.

The invention discloses Pseudomonas sp.HZN6 and application thereof to nicotine degradation. In application, with nicotine as a substrate, an inorganic salt nutrient solution as a reaction medium and Pseudomonas sp.HZN6 as an enzyme, the reaction is carried out for 8-15h under the conditions that temperature is 25-45 DEG C and a Ph value is 5.5-9.0 to ensure that the content of nicotine in the reaction solution is less than 0.1 percent by mass, thus the purpose of degrading nicotine is reached. The Pseudomonas sp.HZN6 can be directly added to a water body and soil to degrade nicotine in the water body and soil, and can be used for safely, efficiently and rapidly degrading residual nicotine on objects such as water and soil. A bactericide containing the Pseudomonas sp.HZN6 is prepared by a simple process with low cost, is convenient to use, and has a good application prospect.

The invention provides an agrobacterium tumefaciens SCUEC1 strain. The collection number of the strain is CCTCC NO:M2014133, and the nucleotide sequence of the strains is as shown in SEQ ID NO. 1. The strains not only can be applied to nicotine degradation, but also can accelerate the maturity of tobacco leaves. Meanwhile, the invention further provides a method for screening and separating the strains. The method comprises the following steps: performing enrichment culture, then performing separation purification, inoculating separated and purified strains to a nicotine culture medium according to a single colony, culturing in a shaking table for 1-3 days at room temperature, measuring the nicotine content of a fermentation broth of the nicotine culture medium, and secondarily screening the strains according to the magnitude of the nicotine degradation rate to obtain the agrobacterium tumefaciens SCUEC1 strains.

The invention relates to a nicotine-degrading bacterium and an application thereof. The nicotine-degrading bacterium is characterized in that the bacterium is named as Arthrobacter histidinolovorans EA-17 according to taxonomy and was collected in China Center for Type Culture Collection on September 15, 2013, and the collection number is CCTCC No. M2013405. A strain is fermented by a 2M3 fermentation tank for production, then mixed with a self-made decomposition agent according to the mass ratio of 1:10, further inoculated into a tobacco straw compost according to the inoculation amount of 1% and fermented for 30-45d, and then straw can be decomposed; compared with traditional compost fermentation, the nicotine degradation rate is improved by 60%-78%, the total nitrogen content is increased by 7%-9%, the total phosphorus content is increased by 0.3%-0.6%, and the total potassium content is increased by 6%-8%.

The present invention provides an Arthrobacter protophormiae SCUEC5 strain, wherein the preservation number is CCTCC NO:M2014404, the nucleotide sequence of the strain is represented by SEQ ID NO.1, and the strain can be applied in nicotine degradation and can further be provided for accelerating tobacco leaf maturation. The present invention further provides a method for screening and separating the strain, wherein the method comprises: carrying out enrichment culture, carrying out separation purification, inoculating the separated and purified single colony into a nicotine culture medium, carrying out shaking culture for 1-3 days at a room temperature, determining the nicotine content in the fermentation broth of the nicotine culture medium, and re-screening the strain according to the nicotine degradation rate so as to obtain the Arthrobacter protophormiae SCUEC5 strain.

The invention relates to a pseudomonas fluorescens preparation and application thereof, and belongs to the field of biotechnology. A preparation method of the pseudomonas fluorescens preparation comprises the following steps of 1, selecting pseudomonas fluorescens 1206 as a production bacterial strain, wherein the pseudomonas fluorescens 1206 has a preservation number of CGMCC NO.3149, 2, carrying out strain culture through inoculating a culture solution with the pseudomonas fluorescens 1206 subjected to inclined plane culture, wherein the culture solution comprises 3% of beef extract, 0.5% of peptone, 0.5% of sodium chloride and the balance distilled water, has a pH value of 7.0 to 7.2 and is put in a shaking bottle, and carrying out culture at a shaking bottle rotate speed of 150rpm at a temperature of 28 to 30 DEG C for 18 hours to obtain a liquid fermented strain, and 3, carrying out fermentation tank fermentation through inoculating a liquid fermentation system in a fermentation tank with the liquid fermented strain, wherein the liquid fermentation system comprises 35g / L of maltose, 4.03g / L of extract yeast, 4.72g / L of ammonium sulfate, 0.4g / L of K2HPO4, 0.075g / L of MgSO4.7H2O, 0.0005g / L of FeSO4.7H2O, 0.015g / L of CaCl2 and 0.1g / L of NaCl, and carrying out fermentation under the conditions of a fermentation temperature of 27 to 29 DEG C, an inoculation amount of 5 to 10%,a fermentation tank stirring speed of 120 to 150rpm, an aeration rate of 5 to 10% and fermentation time of 24 to 48 hours. The pseudomonas fluorescens preparation has a short fermentation period and a high nicotine degradation rate, can improve tobacco quality and can be utilized for degradation of nicotine of tobacco or tobacco waste.

The present invention provides an Ochrobactrum intermedium SCUEC4 strain, wherein the preservation number is CCTCC NO:M2014403, the nucleotide sequence of the strain is represented by SEQ ID NO.1, and the strain can be applied in nicotine degradation and can further be provided for accelerating tobacco leaf maturation. The present invention further provides a method for screening and separating the strain, wherein the method comprises: carrying out enrichment culture, carrying out separation purification, inoculating the separated and purified single colony into a nicotine culture medium, carrying out shaking culture for 1-3 days at a room temperature, determining the nicotine content in the fermentation broth of the nicotine culture medium, and re-screening the strain according to the nicotine degradation rate so as to obtain the Ochrobactrum intermedium SCUEC4 strain.

The present invention relates to a novel tobacco fermenting enzyme preparation production method. The purpose is to solve the technical problems that how improve the quality of the tobacco fermenting product is improved and how the reactivity protection of the tobacco fermenting enzyme preparation is realized in the natural fermenting field of tobacco. The enzyme preparation consists of a glucoseoxidase, a chlorophyl oxidase, a carotenoid oxidase, a protease, and a nicotine-degradation enzyme. Through the cell disruption of fresh leaves, (NH 4) 2 SO 4 is utilized to operate the second fractional precipitation to obtain crude enzyme fluid, an enzyme molecule adopts Ca 2 + and Mg 2 + to operate the metal ion exchange, to accomplish the molecule modification; a macro molecule combination modification is accomplished through adopting 0.01 percent of cane sugar low molecular polymer, thereby prolonging the half life period of theenzyme preparation and obviously improving the high temperature resistant ability. The experimental result employed by the enzyme preparation indicates that the nicotine is decreased by 9.3 percent, the total nitrogen is decreased by 5.7 percent, the protein is decreased by 7.1 percent; cigarette smoke condensates are decreased by 8.4 percent, the tar content is decreased by 5.1 percent, the cigarette smoke nicotine content is decreased by 28.0 percent, and the carbon monoxide is decreased by 1.6 percent. The enzyme preparation is employed when the tobacco leaf is wet for the second time before defolat and redrying.

The invention discloses an Acinetobacter and application thereof. Production bacterial strain liquid is fermented and cultured to form corresponding preparation. The Production bacterial strain is the Acinetobacter (Acinetobacter sp.ND12); and the bacterial strain was preserved in China General Microbiological Culture Collection Center on November 5, 2009, and a preservation number is CGMCC No.3410. The bacterial strain has stronger nicotine degradation capacity and nicotine toxicity resistance capacity, and can be grown at the nicotine concentration of 3.5g / L; and by large-scale culturing of the bacterial strain, the pure nicotine can be degraded by using a whole cell of the bacterial strain as a biocatalyst, and meanwhile the content of the nicotine in tobacco is also reduced. The Acinetobacter and the application thereof have the advantages of capacities of adjusting the content of the nicotine in the tobacco raw material, improving availability of the tobacco, degrading the nicotine in smoke dust, reducing pollution of the nicotine to environment and the like.

The invention provides a Pseudomonas monteilii SCUEC2 strain, which has a preservation number of CCTCC NO:M2014134 and a nucleotide sequence shown as SEQ ID NO.1. The strain not only can be applied to nicotine degradation, but also can accelerate tobacco leaf ripening. The invention also provides a method for screening and separation of the strain. The method comprises the steps of: firstly conducting enrichment culture, then carrying out separation and purification, and finally inoculating the separated and purified strain into a nicotine medium by single colony, performing shake cultivation at room temperature for 1-3d, then measuring the content of nicotine in the fermentation broth of the nicotine medium, and re-screening the strain according to the nicotine degradation rate, thus obtaining the Pseudomonas monteilii SCUEC2 strain.

The invention provides a new strain of nicotine degrading bacteria—Pseudomonas ZUTSKD and its application. Pseudomonas sp. ZUTSKD (Pseudomonas sp. ZUTSKD), preserved in China Center for Type Culture Collection, date of preservation is June 18, 2007, preservation number CCTCCNO: M 207083. The bacteria can degrade nicotine to produce intermediate products such as 2,3'-Bipyridine, cotinine, 3-(3,4-dihydro-2H-pyrrol-5-yl)-Pyridine and 3-Pyridinecarboxylic acid, and can tolerate the highest nicotine With a concentration of 5.5g / L, it can degrade nicotine in waste tobacco powder, and has a certain application prospect in industry.

The invention relates to a novel production method of a tobacco-leaf fermentation enzymatic preparation, which aims at solving the technical problems how to improve tobacco-leaf fermentation quality and realize the activity protection of the tobacco-leaf fermentation enzymatic preparation in the tobacco-leaf spontaneous fermentation field. The enzymatic preparation is composed of glucose oxidase, chlorophyll oxidase, carotenoid oxidase, protease, and nicotine-degradation enzyme. Fresh tobacco leaves are processed through cell disruption, and processed through secondary fractional precipitation by (NH4)2SO4 to obtain crude enzyme liquid; enzyme molecules adopt Ca<2+> and Mg <2+> to act metal ion exchange to complete molecular modification; macromolecule combining modification is completed by adopting sucrose oligomer of 0.01 percent, so that the half-life of the enzymatic preparation can be prolonged, and the heat-resisting ability thereof can be increased markedly. Experimental results which are obtained by using the enzymatic preparation show that the nicotine is reduced 9.3 percent, total nitrogen is reduced 5.7 percent, protein is reduced 7.1 percent, smoke total particulate matter is reduced 8.4 percent, the quantity of the tar is reduced 5.1 percent, the nicotine content in smoke is reduced 28.0 percent, and carbon monoxide is reduced 1.6 percent. The enzymatic preparation is used in the secondary moistening process before threshing and re-drying of the tobacco leaves.

The invention relates to a culture device and a rapid culture method of aerobic granular sludge capable of efficiently degrading nicotine. The device is composed of an SBAR reactor, a numerical control water bath kettle, an air compressor, a gas rotor flow meter, an electronic relay, and a sand core aeration head. The constructed SBAR reactor is used for culturing aerobic granular sludge for degrading nicotine, the temperature of the reactor is about 25 DEG C, and the initial pH value of inlet water is controlled to be about 7.0. In the culture process of the aerobic granular sludge, the nicotine degradation capacity and the COD degradation rate are gradually improved, the average particle size of the sludge is gradually increased, and the biological phase in the reactor also shows an evolution rule from low level to high level. In the process of treating tobacco sheet wastewater by using nicotine degrading bacteria, aerobic granular sludge for efficiently degrading nicotine is cultured and domesticated, and the operation of the SBAR reactor is combined to quickly remove nicotine, so that a good foundation is laid for the research of treating nicotine sheet wastewater by using microorganisms.

The invention relates to a technology of biological degradation of nicotine in tobacco, in particular to a pseudomonad ZCJ bacterial strain applied to nicotine degradation of tobacco and a screening method and an application thereof. The pseudomonad ZCJ bacterial strain (Pseudomonas sp. ZCJ) was preserved in the China Center for Type Culture Collection on March, 30th, 2009 with a preservation number of CCTCC NO. M209064. The invention also discloses screening and application of the bacterial strain. In the invention, nicotine degradation bacterial strain screened from tobacco materials has relatively high biological activity and is applicable to degradation of nicotine in tobacco in a solid state, thus achieving the purpose of the degradation of the nicotine in tobacco.

The invention relates to a culture medium, in particular to a tobacco leaf extract culture medium for cultivating nicotine degradation microorganisms and application thereof and belongs to the technical field of microorganisms. The tobacco leaf extract culture medium provided by the invention mainly comprises tobacco leaf extract. A method for preparing the culture medium comprises the following steps of: mixing tobacco leaf chips and water in a mass volume ratio of 10-80:1; boiling the mixture for 25 to 35 minutes; and performing extraction filtration on the boiled mixture to achieve constant volume so as to obtain the tobacco leaf extract. The invention also provides the application of the tobacco leaf extract culture medium in the aspect of cultivating the nicotine degradation microorganisms. The culture medium is suitable for cultivating various nicotine degradation microorganisms and can obtain high biomass and improve nicotine degradation capability. The culture medium has the advantages of simple preparation, convenient use, low cost and suitability for industrial application.

The invention provides an arthrobacter sp SCUEC3 strain. The preservation number of the strain is CCTCC NO: M2014402. A nucleotide sequence of the strain is shown as SEQ ID NO.1. The strain can be applied to nicotine degradation and can be used for accelerating maturing of tobacco leaves. Meanwhile, the invention also provides a method for screening and separating the strain. The method comprises the following steps: firstly, performing enrichment culture; secondly, performing separation and purification; finally, inoculating the separated and purified strain in a nicotine culture medium according to a single colony, performing shake cultivation at room temperature for 1-3 days, determining the content of nicotine in the fermentation liquor of the nicotine culture medium, and re-screening the strain according to the size of the nicotine degradation rate to obtain the arthrobacter sp SCUEC3 strain.

The invention provides a method for detecting secondary alkaloids produced through nicotine degradation by simulating an actual environment. The method comprises steps as follows: simple sample preparation, determination of an actual environment condition simulation scheme, sampling at multiple points in time, quantitative analysis with gas chromatography-mass spectrometry, data analysis of a measured result and the like. A degradation scheme comprises two major variables, simulation of the real environmental conditions and chemical stability property of nicotine are taken into consideration comprehensively, the operation is simple, and feasibility and scientificity of the scheme are high. The detection method has the advantages of short detection time, high sensitivity and the like. With the application of the technical scheme, the nature of transformation of multi-source nicotine into secondary alkaloids can be taken into consideration comprehensively, a reliable nicotine and secondary alkaloid transformation map is obtained, and scientific support is provided for a tobacco product identification method with the secondary alkaloids as indexes in the tobacco industry.

A nicotine-containing chewing gum piece packed in a wrapping of laminate, where the laminate comprises at least an inner layer facing the chewing gum and a nicotine degradation agent barrier layer of a metal foil. The nicotine-containing chewing gum piece is shaped as an elongate plate having two ends and a thickness and a length, where the length is in the range of 8 to 20 times the thickness. The wrapping has two elongate edges sealed to one another in a fin area extending along the length of the chewing gum piece and two sealed end areas extending beyond the ends of the chewing gum piece.

The invention relates to a nicotine-degrading bacterium and an application thereof. The nicotine-degrading bacterium is characterized in that the bacterium is named as Arthrobacter histidinolovorans EA-17 according to taxonomy and was collected in China Center for Type Culture Collection on September 15, 2013, and the collection number is CCTCC No. M2013405. A strain is fermented by a 2M3 fermentation tank for production, then mixed with a self-made decomposition agent according to the mass ratio of 1:10, further inoculated into a tobacco straw compost according to the inoculation amount of 1% and fermented for 30-45d, and then straw can be decomposed; compared with traditional compost fermentation, the nicotine degradation rate is improved by 60%-78%, the total nitrogen content is increased by 7%-9%, the total phosphorus content is increased by 0.3%-0.6%, and the total potassium content is increased by 6%-8%.

The invention relates to a pseudomonas fluorescens preparation and application thereof, and belongs to the field of biotechnology. A preparation method of the pseudomonas fluorescens preparation comprises the following steps of 1, selecting pseudomonas fluorescens 1206 as a production bacterial strain, wherein the pseudomonas fluorescens 1206 has a preservation number of CGMCC NO.3149, 2, carrying out strain culture through inoculating a culture solution with the pseudomonas fluorescens 1206 subjected to inclined plane culture, wherein the culture solution comprises 3% of beef extract, 0.5% of peptone, 0.5% of sodium chloride and the balance distilled water, has a pH value of 7.0 to 7.2 and is put in a shaking bottle, and carrying out culture at a shaking bottle rotate speed of 150rpm at a temperature of 28 to 30 DEG C for 18 hours to obtain a liquid fermented strain, and 3, carrying out fermentation tank fermentation through inoculating a liquid fermentation system in a fermentation tank with the liquid fermented strain, wherein the liquid fermentation system comprises 35g / L of maltose, 4.03g / L of extract yeast, 4.72g / L of ammonium sulfate, 0.4g / L of K2HPO4, 0.075g / L of MgSO4.7H2O, 0.0005g / L of FeSO4.7H2O, 0.015g / L of CaCl2 and 0.1g / L of NaCl, and carrying out fermentation under the conditions of a fermentation temperature of 27 to 29 DEG C, an inoculation amount of 5 to 10%, a fermentation tank stirring speed of 120 to 150rpm, an aeration rate of 5 to 10% and fermentation time of 24 to 48 hours. The pseudomonas fluorescens preparation has a short fermentation period and a high nicotine degradation rate, can improve tobacco quality and can be utilized for degradation of nicotine of tobacco or tobacco waste.

The invention discloses a construction method and application of pseudomonas JY-Q fructose metabolism gene deleted strain. JY-Q / 5Delta is used as an original strain, the following two isozyme target genes expressing fructose phosphotransferase are determined through genome information analysis of the strain; a knockout vector of a target gene is constructed; the knockout vector is transformed intoan escherichia coli deleted strain WM3064 by heat transfer, which is a donor strain, and pseudomonas JY-Q is used as a receptor bacterium for carrying out parent conjugation; the recombinants exchanged for the first time are screened by utilizing the Kanamycin resistance gene on the vector; and after the obtained strain is subjected to overnight culture expansion, a reverse screening marker SacBgene carried on a carrier is utilized to carry out second exchange of recombinants to obtain the selectively degraded pseudomonas JY-Q fructose metabolic deleted strain. The pseudomonas JY-Q fructosemetabolism deleted strain is applied in nicotine degradation of waste tobacco leaf water extract (TWE).

A nicotine-containing chewing gum piece packed in a wrapping of laminate, where the laminate comprises at least an inner layer facing the chewing gum and a nicotine degradation agent barrier layer of a metal foil. The nicotine-containing chewing gum piece is shaped as an elongate plate having two ends and a thickness and a length, where the length is in the range of 8 to 20 times the thickness. The wrapping has two elongate edges sealed to one another in a fin area extending along the length of the chewing gum piece and two sealed end areas extending beyond the ends of the chewing gum piece.