CN102604853B - Method for degrading vacuum gas oil by applying combination of phanerochete chrysosporium and bacillus - Google Patents

Method for degrading vacuum gas oil by applying combination of phanerochete chrysosporium and bacillus Download PDF

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CN102604853B
CN102604853B CN 201110030280 CN201110030280A CN102604853B CN 102604853 B CN102604853 B CN 102604853B CN 201110030280 CN201110030280 CN 201110030280 CN 201110030280 A CN201110030280 A CN 201110030280A CN 102604853 B CN102604853 B CN 102604853B
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vacuum distillate
degrading
oil
distillate
bacillus
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CN102604853A (en
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杨树林
艾凤祥
赵健烽
徐骏
杨苏平
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China Petroleum and Chemical Corp
Sinopec Yangzi Petrochemical Co Ltd
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China Petroleum and Chemical Corp
Sinopec Yangzi Petrochemical Co Ltd
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Abstract

The invention relates to a method for degrading vacuum gas oil by applying the combination of phanerochete chrysosporium and bacillus. The invention specifically separates bacillus pumilus SWS-0501 with the collection number of CGMCC (China General Microbiological Culture Collection Center) No. 3608. The invention further relates to a method for degrading the vacuum gas oil, which is characterized by combining the phanerochete chrysosporium and the bacillus pumilus to be used for degrading the vacuum gas oil.

Description

Use the method for the flat lead fungi of yellow spore and genus bacillus combination degraded vacuum distillate
Technical field
The invention belongs to petrochemical industry.More specifically, the present invention relates to the biological degradation of vacuum distillate, that is, relate to microorganism vacuum distillate is degraded to lighter component.
Background technology
Oil is very complicated hydro carbons and the mixture of non-hydrocarbons, and crude oil is studied or carried out processing and utilization, all must carry out fractionation to crude oil, to obtain distillate.The distillate of crude oil mainly contains gasoline, kerosene, aviation kerosene, diesel oil, lubricating oil, pitch etc.
Take a broad view of the history of oil Refining Technologies development, the most basic power that promotes the oil Refining Technologies development is how to go out the specification of quality that can satisfy development and the petroleum products of quantitative requirement from the crude production with certain character, composition.In recent years, an important trend of oil Refining Technologies is the light materialization of heavy oil technology, and its background is that the crude oil of World Oil Market obviously became heavy at nearly more than ten years, and the light ends content in the crude oil reduces.Concerning China petroleum refining industry, this problem more has special importance.The Chinese Crude Oils majority is laid particular stress on, and most crude oil contain＞and 500 ℃ vacuum residuum reaches 40%～50%, and Chinese Crude Oils quantitatively also can not satisfy the needs of the national economy of high speed development day by day.Therefore, each state all attaches great importance to the yield that improves light constituent oil in the process of processing fractionation, has adopted the diverse ways such as physics, chemistry that crude oil and vacuum distillate are processed, to improve yield.The most frequently used method has catalytic cracking, hydrocracking, delayed coking, catalytic reforming, alkylation and blending except atmospheric and vacuum distillation.
In recent years, development along with modern biotechnology, scientist utilizes modern bioremediation technology, and the pollution to environment conducts in-depth research to oil, a large amount of results of study prove: there is the microorganism of energy decomposing petroleum hydrocarbon in occurring in nature, can be with the crude oil decomposed, restructuring minute is degraded to lighter component, even is degraded to carbonic acid gas and water.But technique is mainly used in the improvement crude oil pollution at present, solves the environmental problem and the microbial enhanced oil recovery technology that cause because of crude oil pollution, such as Crude oil from CNOOC leakage, soil crude oil pollution and tertiary oil recovery etc.
Summary of the invention
The objective of the invention is to utilize microorganism to the Degradation of vacuum distillate, microorganism not of the same race is made up to process vacuum distillate, the restructuring portions in the vacuum distillate is degraded into lighter component, the open loop of part aromatic hydrocarbon is converted into straight chain hydrocarbon.This technology is applied to the processing of crude oil, can improves the output capacity of light constituent oil, thereby improve the economic benefit of existing crude oil products.
One aspect of the present invention relates to the bacillus pumilus that preserving number is CGMCC No.3608 (Bacillus pumilus) SWS-0501.
The present invention relates to the method for the vacuum distillate of degrading on the other hand, it is characterized in that with preserving number being that the flat lead fungi of yellow spore (Phanerochaete chrysosporium) of ACCC 30553 and bacillus pumilus (Bacillus pumilus) SWS-0501 that preserving number is CGMCC No.3608 combination are used for the degraded vacuum distillate.
In a particular of the inventive method, the ratio of two kinds of bacterial strains of use is 1: 1.
In a particular of the inventive method, degradation condition is that shaking table is cultivated, 35 ℃, and 180rpm, 96 hours.
In a particular of the inventive method, the boiling point of vacuum distillate is 273-451 ℃.
In a particular of the inventive method, the boiling point of vacuum distillate is 299-533 ℃.
In a particular of the inventive method, the boiling point of vacuum distillate is＞533 ℃.
The invention still further relates to preserving number and be the purposes in the degraded vacuum distillate of being combined in of the flat lead fungi of yellow spore (Phanerochaete chrysosporium) of ACCC 30553 and bacillus pumilus (Bacillus pumilus) SWS-0501 that preserving number is CGMCC No.3608.
By the present invention, the vacuum distillate that underpressure distillation is obtained through combination microbiological deterioration after, the carbon chain lengths of hydrocarbon shortens, obtain more short hydrocarbon, be that the change of light constituent cut is many, and a restructuring minute cut tail off, and the output capacity of light constituent oil is improved, the output capacity of heavy component oil reduces, and part has reached the purpose of light materialization of heavy oil.Apply the present invention to have extraordinary application prospect and economic benefit in the modern petrochemical complex.
Description of drawings
Fig. 1 vacuum distillate one gas chromatogram
Fig. 2 makes up the gas chromatogram of vacuum distillate one after the microbiological deterioration
Fig. 3 vacuum distillate three gas chromatograms
Fig. 4 makes up the gas chromatogram of pressing distillate three after the microbiological deterioration
Fig. 5 vacuum distillate four gas chromatograms
Fig. 6 makes up the gas chromatogram of vacuum distillate four after the microbiological deterioration
Embodiment
1, vacuum distillate degradation bacteria and vacuum distillate
Strains A: the flat lead fungi of yellow spore (Phanerochaete chrysosporium), be commonly called as white-rot fungi, available from Chinese agriculture microbial strains preservation administrative center (Agricultural Culture Collection of China ACCC), preserving number is 30553.
The screening of bacterial strain B:
Take from former greasy dirt and oily sludge 2～3 grams of raising sub-refinery, place the 250mL beaker, add distilled water 150mL, place on the magnetic stirring apparatus, temperature is 20～30 ℃ of room temperatures, stirring velocity 180rpm, then stir process 2.5～3 hours uses the sterilized double layer filtered through gauze, remove upper strata floating matter and lower sediment thing, supernatant liquor is as the bacteria suspension that separates the purpose bacterial strain, and is placed in the aseptic triangular flask, wraps up for subsequent use.Whole operating process is finished at aseptic operating platform.Get respectively l mL bacteria suspension and cultivated 96 hours in the dull and stereotyped coating of crude oil, picking colony is to beef extract-peptone solid medium (prescription (L -1): extractum carnis 5g, peptone 10g, NaCl 5g, agar 18g) upper purifying three times, with the bacterial strain behind the purifying at the dull and stereotyped multiple sieve of crude oil, 30 ℃, cultivated 72 hours, bacterial strain that can continued growth is to utilize the also purpose bacterial strain of degrading crude oil.
The crude oil flat board was composed as follows when crude oil screened and sieves again: add crude oil 0.5% (w/v), agar 4% (w/v) in the ion substratum.
Consisting of of ion substratum: NaNO 31g, KH 2PO 42g, K 2HPO 43H 2O1g, MgSO 47H 2O 0.5g, KCl 0.1g, CaCl 20.01g, FeSO 40.01g, liquid microelement 0.025mL.Liquid microelement forms: boric acid 0.25g, cupric sulfate pentahydrate 0.25g, manganous sulfate 0.5g, Sodium orthomolybdate 0.05g, zinc sulfate 0.7g, water 1000mL.Screen altogether 78 strains of oil degradation bacterium, conservation is for subsequent use after identifying.
Choose a strain microorganism of screening, be designated as SWS-0501.Its Physiology and biochemistry experimental result is as follows
Annotate :+expression is positive;-expression is negative;---the expression undetermined.
The microscope morphological observation
Bacterial strain 0501 is Gram-positive, and the microscopic examination bacterial strain is rod-short, has gemma, bacterium colony to be khaki color, surface drying, fold is arranged;
According to the above results, this bacterial strain called after bacillus pumilus SWS-0501 (hereinafter referred bacterial strain B), it is preserved in (No. 3, No. 3 institutes in BeiChen West Road, Chaoyang District, BeiJing City of Chinese microorganism strains preservation management committee common micro-organisms center (CGMCC) on January 25th, 2010, Institute of Microorganism, Academia Sinica, postcode 100101), preserving number is CGMCC No.3608.
Vacuum distillate is that four kinds of vacuum distillates that sub-petrochemical industry lu ning pipeline transportation of crude oil carries out underpressure distillation are raised by Sinopec Group.Vacuum distillate one is liquid, and vacuum distillate two, three and four is condensed state.The boiling spread of each cut is vacuum distillate one: 273-451 ℃; Vacuum distillate two: 311-489 ℃; Vacuum distillate three: 299-553 ℃; Vacuum distillate four:＞553.
2, vacuum distillate degradation method general introduction
With two strain microorganisms as degraded vacuum distillate compatibility bacterium.Strains A is inoculated in adds rich PDA (prescription is: peeling potato 200g; Glucose 20g; KH 2PO 43g, MgSO 47H 2O1.5g; Water 1000mL; Agar 18g; PH nature) on the flat board, 35 ℃ leave standstill and cultivated 48 hours, after all covering with white spore on the flat board, spore inoculating 3 encircled add rich PD liquid nutrient medium in 50mL (prescription is: peeling potato 200g; Glucose 20g; KH 2PO 43g, MgSO 47H 2O 1.5g; Water 1000mL; PH nature) amplification cultivation, culture condition is 35 ℃, 180rpm, 96 hours, when treating strains A to grow into diameter being bacterium ball about 5mm, bacterium liquid is sub-packed in the 40mL centrifuge tube, 10000rpm is centrifugal, removes supernatant liquor, and thalline is for subsequent use.Bacterial strain B is inoculated in beef extract-peptone liquid nutrient medium (prescription (L -1): extractum carnis 5g; Peptone 10g; NaCl 5g; Water 1000mL.) in, the shaking table culture condition is: 30 ℃, and 180rpm, 48 hours.Bacterium liquid is sub-packed in the 40mL centrifuge tube, and 8000rpm is centrifugal, removes supernatant liquor, and thalline is for subsequent use.
Thalline is transferred to (the reduced pressure distillate oil-contg is 0.5g) in the 50mL vacuum distillate liquid nutrient medium, and degradation condition is that shaking table is cultivated, and culture condition is 35 ℃, 180rpm, 96 hours.
3, gas chromatographic analysis
Fermented liquid after the degraded is centrifugal with the speed of 8000rpm, remove thalline, supernatant liquor adopts dichloromethane extraction four times, 0.22 μ m membrane filtration behind the evaporating solvent, adopts GC to analyze, adopt external standard method to determine the composition of the hydrocarbon in the vacuum distillate, it is quantitative that the employing normalization method carries out integration.Analyze the carbon chain lengths of finding hydrocarbon in the vacuum distillate and shorten, mink cell focus partly changes lightweight oil into.
Embodiment one:
1, the cultivation of vacuum distillate degradation bacteria
Strains A is inoculated in adds rich PDA (prescription is: peeling potato 200g; Glucose 20g; KH 2PO 43g, MgSO 47H 2O 1.5g; Water 1000mL; Agar 18g; PH nature) on the flat board, 35 ℃ leave standstill and cultivated 48 hours, after all covering with white spore on the flat board, spore inoculating 3 encircled add rich PD liquid nutrient medium in 50mL (prescription is: peeling potato 200g; Glucose 20g; KH 2PO 43g, MgSO 47H 2O 1.5g; Water 1000mL; PH nature) amplification cultivation, culture condition are that shaking table is cultivated, 35 ℃, and 180rpm, 96 hours, when treating strains A to grow into diameter being bacterium ball about 5mm, bacterium liquid is sub-packed in the 40mL centrifuge tube, 10000rpm is centrifugal, removes supernatant liquor, and thalline is for subsequent use.
Bacterial strain B streak inoculation is in beef extract-peptone solid medium (its prescription (L -1): extractum carnis 5g; Peptone 10g; NaCl 5g; Agar 18g) on, cultivated 24 hours for 30 ℃, inoculation 3 rings are to 150mL beef extract-peptone liquid nutrient medium (its prescription (L -1): extractum carnis 5g; Peptone 10g; NaCl 5g; Agar 18g, water 1000mL) middle amplification cultivation, culture condition is that shaking table is cultivated, 30 ℃, 180rpm, 48 hours, measure its optical density(OD), bacteria concentration reaches 10 8Individual mL -1The time, bacterium liquid is sub-packed in the 40mL centrifuge tube, 10000rpm is centrifugal, removes supernatant liquor, and thalline is for subsequent use.
2, vacuum distillate degraded
Adopt the decrement method accurately to take by weighing vacuum distillate one 0.5g and add (NaNO in the 50mL ion salt substratum 31gl -1, KH 2PO 42gl -1, K 2HPO 43H 2O 1gl -1, MgSO 47H 2O0.5gl -1, KCl 0.1gl -1, CaCl 20.01gl -1, FeSO 40.01gl -1, liquid microelement 0.025mL.Liquid microelement forms: boric acid 0.25gl -1, cupric sulfate pentahydrate 0.25gl -1, manganous sulfate 0.5gl -1, Sodium orthomolybdate 0.05gl -1, zinc sulfate 0.7gl -1), sterilization.
Adopt sterile distilled water that two kinds of bacterium are transferred to the ion salt substratum that contains vacuum distillate, inoculative proportion is 1: 1, places on the shaking table and cultivates, and culture condition is 35 ℃, 180rpm, 96 hours.
3, gas chromatographic analysis
Fermented liquid after the degraded is sub-packed in the 40mL sterilization centrifuge tube, speed with 8000rpm is centrifugal, remove thalline, supernatant liquor and oil residues thereof adopt dichloromethane extraction four times, total extraction agent add-on 20mL, and extraction liquid is through 0.22 μ m membrane filtration, getting 1mL is sub-packed in the 1.5mL centrifuge tube, place the air-dry methylene chloride of stink cupboard, remaining vacuum distillate adopts the dissolving of 60 μ l methylene dichloride, carries out GC and analyzes.The GC test condition is: instrument: Agilent 5890; Detector: FID; Post type: HP-5; Sensitivity: 2; Post specification: 0.25 μ m * 30m; Hydrogen: 0.14MPa; Carrier gas type: N 2Air: 0.14MPa; Sampler temperature: 340 ℃; Detector temperature: 340 ℃; Sample size: 0.4 μ l.Temperature programming: 80 ℃ of initial temperatures: initial retention time 5 minutes; 4 ℃/minute of temperature rise rates; 320 ℃ of final temperatures kept 10 minutes.Adopt external standard method to determine the composition of the hydrocarbon in the vacuum distillate, it is quantitative that the employing normalization method carries out integration.External standard accepted standard material is available from ChemService, Petrochemical Calibration Mixture#1 (ASTM D2887), compositions of mixtures is C6, C7, C8, C9, C10, C11, C12, C14, C16, C18, C20, C24, C28, C32, C36, C40, C44, and content is respectively 6%, 6%, 8%, 8%, 12%, 12%, 12%, 12%, 10%, 5%, 2%, 2%, 1%, 1%, 1%, 1%, 1%.
The gas-chromatography spectrogram is seen Fig. 1 and Fig. 2 before and after vacuum distillate one degraded.
Variation has occured in content in the straight-chain paraffin in the analysis discovery vacuum distillate one, C22 and C23 content reduce, and the content of C18, C20 and C21 increases, and the subtle change that the content of other alkane also occurs illustrates that microorganism becomes short alkane with the alkane degradation of long-chain.And other short chain alkanes content does not reduce.Generally, vacuum distillate one is through after making up microbiological deterioration, and light hydrocarbon component increases.
Embodiment two:
1, the cultivation of vacuum distillate degradation bacteria
With embodiment one.
2, vacuum distillate degraded
The vacuum distillate that adopts is reduced pressure distillate three, and other are with embodiment one.
3, gas chromatographic analysis
Other are with embodiment one.Fig. 3 and Fig. 4 are seen in gas chromatographic analysis after the degraded.
Analyze and find that C33～C35 has obvious degraded in various degree in the vacuum distillate, other hydro carbons also have degraded in various degree, and the amount of short hydrocarbon does not reduce.And increased new hydro carbons composition between C24～C26.
Embodiment three:
1, the cultivation of vacuum distillate degradation bacteria
With embodiment one.
2, vacuum distillate degraded
The vacuum distillate that adopts is vacuum distillate four, and other are with embodiment one.
3, gas chromatographic analysis
Other are with embodiment one.Fig. 5 and Fig. 6 are seen in gas chromatographic analysis after the degraded.
Analyze and find the following generally content increase of straight-chain paraffin of C29 in the vacuum distillate four, and the component kind between C31 and the C36 increases, and straight-chain paraffin C36 content reduces at most, C31 content increases at most.Vacuum distillate four is described through after the microbiological deterioration, lightweight oil content increases.

Claims (7)

1. preserving number is bacillus pumilus (Bacillus pumilus) SWS-0501 of CGMCC No.3608.
2. the method for degraded vacuum distillate is characterized in that with preserving number being that the flat lead fungi of yellow spore (Phanerochaete chrysosporium) of ACCC 30553 and bacillus pumilus (Bacillus pumilus) SWS-0501 that preserving number is CGMCC No.3608 combination are used for the distillate of degrading.
3. the method for claim 2, the ratio of use therein two kinds of bacterial strains is 1:1.
4. claim 2 or 3 method, wherein degradation condition is that shaking table is cultivated, 35 ℃, 180rpm, 96 hours.
5. claim 2 or 3 method, wherein the boiling point of vacuum distillate is 273-451 ℃.
6. claim 2 or 3 method, wherein the boiling point of vacuum distillate is 299-533 ℃.
7. claim 2 or 3 method, wherein the boiling point of vacuum distillate is〉533 ℃.
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CN1252248C (en) * 2003-10-24 2006-04-19 中国石油化工股份有限公司 Bacillus subtilis and its application
CN1236054C (en) * 2004-05-17 2006-01-11 大庆油田有限责任公司 Bacterium for degrading petroleum and it use
CN1236053C (en) * 2004-05-17 2006-01-11 大庆油田有限责任公司 Bacterium for degrding petroleum and its use
CN100487108C (en) * 2007-03-16 2009-05-13 山东省科学院生物研究所 Solid microbe agent for degrading petroleum pollution, and petroleum products, and preparation method
CN101560483B (en) * 2009-05-31 2010-11-17 南京农业大学 Lipopeptide-producing bacillus pumilus and application thereof

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