CN108753856A - A method of it is accumulated based on molasses alcohol waste mash culture microalgae grease - Google Patents
A method of it is accumulated based on molasses alcohol waste mash culture microalgae grease Download PDFInfo
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- 239000004519 grease Substances 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title claims abstract description 16
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 title claims abstract description 15
- 239000002699 waste material Substances 0.000 title claims abstract description 14
- 241000195493 Cryptophyta Species 0.000 claims abstract description 36
- 239000003225 biodiesel Substances 0.000 claims abstract description 28
- 239000001963 growth medium Substances 0.000 claims abstract description 21
- 230000005526 G1 to G0 transition Effects 0.000 claims abstract description 9
- 230000006698 induction Effects 0.000 claims abstract description 9
- 239000007640 basal medium Substances 0.000 claims abstract description 7
- 238000004519 manufacturing process Methods 0.000 claims abstract description 7
- 239000002609 medium Substances 0.000 claims abstract description 6
- 239000012452 mother liquor Substances 0.000 claims abstract description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 18
- 238000005286 illumination Methods 0.000 claims description 17
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 12
- 238000000605 extraction Methods 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 claims description 6
- 239000012141 concentrate Substances 0.000 claims description 6
- 239000012153 distilled water Substances 0.000 claims description 6
- 230000032050 esterification Effects 0.000 claims description 6
- 238000005886 esterification reaction Methods 0.000 claims description 6
- WCYAALZQFZMMOM-UHFFFAOYSA-N methanol;sulfuric acid Chemical compound OC.OS(O)(=O)=O WCYAALZQFZMMOM-UHFFFAOYSA-N 0.000 claims description 6
- 239000011259 mixed solution Substances 0.000 claims description 6
- 239000003960 organic solvent Substances 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 239000004576 sand Substances 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 239000000243 solution Substances 0.000 claims description 4
- 229910052564 epsomite Inorganic materials 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims 2
- 238000000227 grinding Methods 0.000 claims 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims 1
- 239000010453 quartz Substances 0.000 claims 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims 1
- 238000005516 engineering process Methods 0.000 abstract description 2
- 238000011534 incubation Methods 0.000 abstract 1
- 238000004904 shortening Methods 0.000 abstract 1
- 239000002028 Biomass Substances 0.000 description 15
- 239000012074 organic phase Substances 0.000 description 5
- 239000011044 quartzite Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 238000003912 environmental pollution Methods 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 235000019737 Animal fat Nutrition 0.000 description 1
- 239000004215 Carbon black (E152) Substances 0.000 description 1
- CZMRCDWAGMRECN-UHFFFAOYSA-N Rohrzucker Natural products OCC1OC(CO)(OC2OC(CO)C(O)C(O)C2O)C(O)C1O CZMRCDWAGMRECN-UHFFFAOYSA-N 0.000 description 1
- 240000000111 Saccharum officinarum Species 0.000 description 1
- 235000007201 Saccharum officinarum Nutrition 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000005791 algae growth Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000002551 biofuel Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000002485 combustion reaction Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000002283 diesel fuel Substances 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 235000019441 ethanol Nutrition 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000005431 greenhouse gas Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000011169 microbiological contamination Methods 0.000 description 1
- 239000002366 mineral element Substances 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 230000036284 oxygen consumption Effects 0.000 description 1
- 230000032696 parturition Effects 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 238000005809 transesterification reaction Methods 0.000 description 1
- 235000019871 vegetable fat Nutrition 0.000 description 1
Classifications
-
- 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
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6436—Fatty acid esters
- C12P7/649—Biodiesel, i.e. fatty acid alkyl esters
-
- 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
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention discloses a kind of method accumulated using molasses alcohol waste mash culture microalgae grease, belongs to biodiesel technology field.The method of the invention includes the following steps：Useless mash is diluted, as the minimal medium of single needle algae, MgSO is added into basal medium4·7H2O, adjustment pH value are 6.8-7.1, and culture medium is prepared；Above-mentioned culture medium is sterilized under high-temperature and high-pressure conditions, accesses oil-producing single needle algae, algae cell density is 1 × 106‑1.5×106A/ml, then addition prepare epiphysin mother liquor with absolute ethyl alcohol, and epiphysin mother liquor is added in the induction algae solution diluted makes a concentration of the 10 of final epiphysin‑4μm ol/L-1 μm of ol/L, and algae solution is placed under intense light irradiation and is cultivated；The microalgae of culture to stationary phase is enriched with, grease is extracted, biodiesel is prepared.Present invention incubation time that is simple and practicable, can substantially shortening oil-producing single needle algae reduces production cost, and dramatically increases grease yield, to substantially increase the efficiency that microalgae produces biodiesel.
Description
Technical field
The present invention relates to a kind of methods accumulated using molasses alcohol waste mash culture microalgae grease, belong to biodiesel skill
Art field.
Background technology
Biodiesel refers to by transesterification or thermochemical processes with vegetable fat, microbial grease, animal fat, useless meal
Drink oil etc. can replace the reproducibility diesel fuel of petrifaction diesel made of raw material.Compared with common diesel, it have environmental protection, can
Regeneration, degradable, post-combustion pollution object discharge the features such as low, greenhouse gas emission is low.
Microalgae produces the raw material of bio-fuel as terrestrial plant is substituted, can be effectively sharp because it can carry out photosynthesis
With solar energy by water, CO2It is converted into organic matter with inorganic salts.Microalgae is high with photosynthetic utilization rate, the speed of growth is fast, to growth ring
Border requires low and ergastic substances（Grease, hydrocarbon）The advantages that content is high has been widely studied.Therefore, biodiesel is produced using microalgae
With vast potential for future development.Present microalgae mainly uses light autotrophy culture.But light autotrophy training mode, which has, makes frustule give birth to
Long period is long, low yield, easy microbiological contamination and harvesting of high cost the features such as.And and because have illumination and organic carbon source under the conditions of supporting,
Biomass can be largely accumulated, cell growth cycle is shortened.But simultaneous foster Biomass yield and high-quality yield are relatively low therefore simultaneous
It is unsuitable for accumulating grease as supporting.
Molasses alcohol waste mash is sugarcane or beet-sugar factory during refined sugar product, and the by-product molasses of gained pass through
The useless mash discharged after fermentation and distillation, chemical oxygen consumption (COC) (COD) reach 40000-50000mg/L, BOD (BOD)
Up to 20000-30000mg/L, ammonia nitrogen 1500-2500mg/L, wherein also contain there are many amino acid, protein, carbohydrate and Na, K,
The mineral elements such as Ca, Mg, P, solid content 8%-10%.How cost-effective improvement is carried out to alcohol waste mash, until
The present is still the problem that sugar plant pollution is administered.And use useless mash as minimal medium culture microalgae and produce biodiesel, no
It can only solve the problems, such as environmental pollution treatment, moreover it is possible to bring economic benefit for enterprise, can accomplish waste utilization completely, this will be
Sugar enterprise of Yunnan Province brings good foreground.
Invention content
The object of the present invention is to provide a kind of method for giving birth to oil and fat accumulation using molasses alcohol waste mash culture microalgae, the party
Method process is simple, it is preferred that emphasis is culture, production cost is low, specifically includes following steps：
（1）Prepare culture medium：Useless mash is diluted, as the minimal medium of single needle algae, is added into basal medium
MgSO4·7H2O, adjustment pH value are 6.8-7.1, and culture medium is prepared；
（2）Oil-producing microalgae culture：Above-mentioned culture medium is sterilized under high-temperature and high-pressure conditions, accesses oil-producing single needle algae, frustule is close
Degree is 1 × 106-1.5×106A/ml, then addition prepare epiphysin mother liquor with absolute ethyl alcohol, epiphysin mother liquor are added to
Make a concentration of the 10 of final epiphysin in induction algae solution through having diluted-4μm ol/L-1 μm of ol/L, and algae solution is placed in strong light
According to lower culture；
（3）Prepare biodiesel：The microalgae of culture to stationary phase is enriched with, grease is extracted, biodiesel is prepared.
Preferably, step of the present invention（1）In give up mash extension rate be 1550-1650 times.
Preferably, MgSO of the present invention4·7H2The addition of O is 176.5-187.9mg/L.
Preferably, step of the present invention（2）Middle condition of culture is：Illumination cultivation temperature is 23-26 DEG C, and intensity of illumination is
2500-4000lux, shaking speed 130-160r/min.
Preferably, the step of the present invention（3）Middle preparation biodiesel specific method is：By medium centrifugal richness
Collection, centrifugal condition are to centrifuge 10min under 3500r/min speed conditions, and be lyophilized after washing 2 times repeatedly with distilled water；Addition is frozen
It is 2 that volume ratio is used after the quartzite sand grind 20min of 2 times of weight of dry algae powder:1 chloroform-methanol carries out grease extraction, organic solvent
It repeats to collect organic phase concentrate after extracting 3 times, the mixed solution of 3% sulfuric acid-methanol is added in the grease extracted（Sulfuric acid with
The volume ratio of methanol is 3:97）It carries out esterification and biodiesel is prepared.
The beneficial effects of the invention are as follows：
（1）The present invention can save carbon source and nitrogen source and other trace elements, improve the utilization rate and conversion ratio of sugar, reduce
Cost needed for production；Useless mash realizes zero-emission, is conducive to environmental protection；The processing of environmental pollution and the culture of microalgae are combined
Together, energy saving, turn waste into wealth.
（2）For the present invention compared with useless mash individually culture, fat content and biomass have promotion by a relatively large margin；It reaches
Maximum biomass when stationary phase improves 11.8-42.3% compared with control, reaches 0.856-1.218g/L, and fat content improves
27.8-65.6%, has reached 53.08-68.77%.
（3）Oil-producing microalgae can be grown in useless mash culture medium well in the present invention, not only solved environmental pollution and asked
Topic, and convert pollutant to reproducible bioenergy；Microalgae can not only be grown well in useless mash, and can also
Promote the increase of oil-producing microalgae fat content；Mechanism of action that may be present is that the nitrogen source, carbon source inside useless mash can be by microalgaes
It absorbs and utilizes well, be also likely to be present the acting factor for promoting micro algae growth inside useless mash, need further to probe into.
Description of the drawings
Fig. 1 is the interpretation of result figure of embodiment 3.
Specific implementation mode
The present invention is described in detail With reference to embodiment, but protection scope of the present invention is not limited to institute
State content.
Reference examples 1
（1）Prepare culture medium：By 1600 times of useless mash gradient dilution, as single needle algaeMonoraphidiumSp. FXY-10 bases
Then MgSO is added in basal culture medium into basal medium4·7H2O：Dosage 182.2mg/L；Adjustment pH value is 6.8-7.0；
（2）Oil-producing microalgae culture：High pressure-temperature sterilizing 20min, accesses oil-producing single needle algae, and algae cell density is controlled 106A/
Epiphysin is added into culture medium and so that a concentration of 0 μm of ol/L of final epiphysin, illumination cultivation temperature are 25 DEG C by ml, illumination
Intensity is 3500lux, shaking speed 150r/min, carries out illumination shaking flask culture；
（3）Prepare biodiesel：Will culture to stationary phase microalgae be enriched with, centrifugal condition be 3500r/min speed conditions under from
Heart 10min, and be lyophilized after washing 2 times repeatedly with distilled water；It is used after adding the quartzite sand grind 20min of 2 times of weight of freeze-dried algae powder
Volume ratio is 2:1 chloroform-methanol carries out grease extraction, and organic solvent repeats to collect organic phase concentrate, extraction after extracting 3 times
The mixed solution of 3% sulfuric acid-methanol is added in obtained grease（The volume ratio of sulfuric acid and methanol is 3:97）Esterification is carried out to be prepared into
To biodiesel.
Fat content has reached 41.54% under the induction of 0 μm of ol/L epiphysin, and biomass has reached 0.571g/L.
Embodiment 1
（1）Prepare culture medium：By 1600 times of useless mash gradient dilution, as single needle algaeMonoraphidiumSp. FXY-10 bases
Then MgSO is added in basal culture medium into basal medium4·7H2O：Dosage 182.2mg/L；Adjustment pH value is 6.8-7.0；
（2）Oil-producing microalgae culture：High pressure-temperature sterilizing 20min, accesses oil-producing single needle algae, and algae cell density is controlled 106A/
Epiphysin is added into culture medium and so that a concentration of 1 μm of ol/L of final epiphysin, illumination cultivation temperature are 25 DEG C by ml, illumination
Intensity is 3500lux, shaking speed 150r/min, carries out illumination shaking flask culture.
（3）Prepare biodiesel：The microalgae of culture to stationary phase is enriched with, centrifugal condition is 3500r/min speed conditions
Lower centrifugation 10min, and be lyophilized after washing 2 times repeatedly with distilled water；Add the quartzite sand grind 20min of 2 times of weight of freeze-dried algae powder
It is 2 to use volume ratio afterwards:1 chloroform-methanol carries out grease extraction, and organic solvent repeats to collect organic phase concentrate after extracting 3 times,
Extract the mixed solution that 3% sulfuric acid-methanol is added in obtained grease（The volume ratio of sulfuric acid and methanol is 3:97）The esterification system of progress
It is standby to obtain biodiesel.
The oil-producing single needle algae fat content of the present embodiment culture has reached 62.46%, and biomass has reached 1.075g/L.
Embodiment 2
（1）Prepare culture medium：By 1550 times of useless mash gradient dilution, as single needle algaeMonoraphidiumSp. FXY-10 bases
Then MgSO is added in basal culture medium into basal medium4·7H2O：Dosage 176.5mg/L；Adjustment pH value is 6.8-7.0；
（2）Oil-producing microalgae culture：High pressure-temperature sterilizing 20min, accesses oil-producing single needle algae, and algae cell density is controlled 1.2 × 106
A/ml, into culture medium, addition epiphysin makes final epiphysin a concentration of 10-3μm ol/L, illumination cultivation temperature are 23 DEG C,
Intensity of illumination is 4000lux, shaking speed 130r/min, carries out illumination shaking flask culture.
（3）Prepare biodiesel：The microalgae of culture to stationary phase is enriched with, centrifugal condition is 3500r/min speed conditions
Lower centrifugation 10min, and be lyophilized after washing 2 times repeatedly with distilled water；Add the quartzite sand grind 20min of 2 times of weight of freeze-dried algae powder
It is 2 to use volume ratio afterwards:1 chloroform-methanol carries out grease extraction, and organic solvent repeats to collect organic phase concentrate after extracting 3 times,
Extract the mixed solution that 3% sulfuric acid-methanol is added in obtained grease（The volume ratio of sulfuric acid and methanol is 3:97）The esterification system of progress
It is standby to obtain biodiesel.
The oil-producing single needle algae fat content of the present embodiment culture has reached 68.77%, and biomass has reached 1.218g/L.
Embodiment 3
（1）Prepare culture medium：By 1650 times of useless mash gradient dilution, as single needle algaeMonoraphidiumSp. FXY-10 bases
Then MgSO is added in basal culture medium into basal medium4·7H2O：Dosage 187.5mg/L；Adjustment pH value is 6.8-7.0；
（2）Oil-producing microalgae culture：High pressure-temperature sterilizing 20min, accesses oil-producing single needle algae, and algae cell density is controlled 1.5 × 106
A/ml, into culture medium, addition epiphysin makes final epiphysin a concentration of 10-4μm ol/L, illumination cultivation temperature are 26 DEG C,
Intensity of illumination is 2500lux, shaking speed 160r/min, carries out illumination shaking flask culture.
（3）Prepare biodiesel：The microalgae of culture to stationary phase is enriched with, centrifugal condition is 3500r/min speed conditions
Lower centrifugation 10min, and be lyophilized after washing 2 times repeatedly with distilled water；Add the quartzite sand grind 20min of 2 times of weight of freeze-dried algae powder
It is 2 to use volume ratio afterwards:1 chloroform-methanol carries out grease extraction, and organic solvent repeats to collect organic phase concentrate after extracting 3 times,
Extract the mixed solution that 3% sulfuric acid-methanol is added in obtained grease（The volume ratio of sulfuric acid and methanol is 3:97）The esterification system of progress
It is standby to obtain biodiesel.
The oil-producing single needle algae fat content of the present embodiment culture has reached 53.08%, and biomass has reached 1.119g/L.
Under the same conditions, epiphysin concentration is set to 1 μm of ol/L, 10-1μmol/L、10-2μmol/L、10-3μ
mol/L、10-4μm ol/L, it is as a result as follows：
Fat content has reached 62.46% under the induction of 1 μm of ol/L epiphysin, and biomass has reached 1.075g/L.
10-1Fat content has reached 63.02% under the induction of μm ol/L epiphysins, and biomass has reached 0.957g/L.
10-2Fat content has reached 67.00% under the induction of μm ol/L epiphysins, and biomass has reached 1.061g/L.
10-3Fat content has reached 68.77% under the induction of μm ol/L epiphysins, and biomass has reached 1.218g/L.
10-4Fat content has reached 53.08% under the induction of μm ol/L epiphysins, and biomass has reached 1.119g/L.
The results are shown in Figure 1, and the present invention is compared with useless mash individually culture as seen from the figure, fat content and biomass
There is promotion by a relatively large margin；Maximum biomass when reaching stationary phase improves 11.8-42.3% compared with control, reaches
0.856-1.218g/L, fat content improve 27.8-65.6%, have reached 53.08-68.77%.
Claims (5)
1. a kind of method producing biodiesel using molasses alcohol waste mash culture single needle algae, which is characterized in that including following
Step：
（1）Prepare culture medium：Useless mash is diluted, as the minimal medium of single needle algae, is added into basal medium
MgSO4·7H2O, adjustment pH value are 6.8-7.1, and culture medium is prepared；
（2）Oil-producing microalgae culture：Above-mentioned culture medium is sterilized under high-temperature and high-pressure conditions, accesses oil-producing single needle algae, frustule is close
Degree is 1 × 106-1.5×106A/ml, then addition prepare epiphysin mother liquor with absolute ethyl alcohol, epiphysin mother liquor are added to
Make a concentration of the 10 of final epiphysin in induction algae solution through having diluted-4μm ol/L-1 μm of ol/L, and algae solution is placed in strong light
According to lower culture；
（3）Prepare biodiesel：The microalgae of culture to stationary phase is enriched with, grease is extracted, biodiesel is prepared.
2. the method for utilizing molasses alcohol waste mash culture single needle algae production biodiesel according to claim 1, feature
It is：Step（1）In give up mash extension rate be 1550-1650 times.
3. the method for utilizing molasses alcohol waste mash culture single needle algae production biodiesel according to claim 1, feature
It is： MgSO4·7H2The addition of O is 176.5-187.9mg/L.
4. the method for utilizing molasses alcohol waste mash culture single needle algae production biodiesel according to claim 1, feature
It is：The step（2）Middle condition of culture is：Illumination cultivation temperature is 23-26 DEG C, and intensity of illumination 2500-4000lux shakes
Bed rotating speed is 130-160r/min.
5. the method for utilizing molasses alcohol waste mash culture single needle algae production biodiesel according to claim 1, feature
It is：The step（3）Middle preparation biodiesel specific method is：Medium centrifugal is enriched with, centrifugal condition is 3500 r/
10min is centrifuged under min speed conditions, and is lyophilized after washing 2 times repeatedly with distilled water；Add the quartz of 2 times of weight of freeze-dried algae powder
It is 2 that volume ratio is used after sand grinding 20min:1 chloroform-methanol carries out grease extraction, and organic solvent, which repeats to collect after extracting 3 times, to be had
The mixed solution of 3% sulfuric acid-methanol is added in machine phase concentrate, the grease extracted（The volume ratio of sulfuric acid and methanol is 3:97）
It carries out esterification and biodiesel is prepared.
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| CN110218652A (en) * | 2019-05-27 | 2019-09-10 | 昆明理工大学 | A method of promoting micro algae growth and oil and fat accumulation in BG-11 culture medium |
Citations (3)
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| CN105039437A (en) * | 2015-07-01 | 2015-11-11 | 昆明理工大学 | Method using molasses, alcohol, and waste undecanted wine to culture monoraphidium to produce diesel oil |
| CN105803010A (en) * | 2016-03-30 | 2016-07-27 | 昆明理工大学 | Method for oil accumulation based on heterotrophic microalgae |
| CN107475171A (en) * | 2017-08-15 | 2017-12-15 | 昆明理工大学 | Application of the epiphysin in oil-producing microalgae fat content is improved |
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| CN105039437A (en) * | 2015-07-01 | 2015-11-11 | 昆明理工大学 | Method using molasses, alcohol, and waste undecanted wine to culture monoraphidium to produce diesel oil |
| CN105803010A (en) * | 2016-03-30 | 2016-07-27 | 昆明理工大学 | Method for oil accumulation based on heterotrophic microalgae |
| CN107475171A (en) * | 2017-08-15 | 2017-12-15 | 昆明理工大学 | Application of the epiphysin in oil-producing microalgae fat content is improved |
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| CN110218652A (en) * | 2019-05-27 | 2019-09-10 | 昆明理工大学 | A method of promoting micro algae growth and oil and fat accumulation in BG-11 culture medium |
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