CN104561147B - A kind of sterilizing methods of liquid alkane - Google Patents
A kind of sterilizing methods of liquid alkane Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
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- 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
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Abstract
The invention discloses a kind of sterilizing methods of liquid alkane, including herein below：For the liquid alkane of bioconversion, handled through microwave demulsifying, alkane and water stratification, add bacteriostatic agent, carry out microwave sterilization processing, the liquid alkane sterilized.Microwave demulsifying processing condition be：Microwave frequency is 300~3000MHz, and Microwave Power Density is 100W/L~10kW/L, and treatment temperature is 50 ~ 90 DEG C, and processing time is 10 ~ 30 minutes.Microwave sterilization processing condition be：Microwave frequency is 300~3000MHz, and Microwave Power Density is 20~100W/L, and processing time is 5~10 minutes.The problem of the inventive method solves alkane loss present in conventional steam sterilizing and high microbiological contamination risk, has broad application prospects.
Description
Technical field
The invention belongs to biological chemical field, more particularly to a kind of sterilizing of the liquid alkane for bioconversion substrate
Method.
Background technology
Alkane, i.e. saturated hydrocarbons（Saturated group）It is the chain hydrocarbon of only carbon-carbon single bond and C-H bond, is simplest
One class organic compound.Alkane property is stablized relatively, is not susceptible to reaction, is industrially typically used directly as fuel.Liquid
State alkane refers to the alkane containing 5~16 carbon atoms, and liquid is typically exhibited at ambient temperature.With the development of biotechnology,
Liquid alkane is widely used in fermentation arts as carbon source, and the fermentation of SL-AH is such as produced with 12 carbon n-alkanes
Technique.
In fermentation production process, medium component is needed by disinfecting action, so as to ensure the strain list of fermentation system
First, stably.Liquid alkane is used usually as carbon source and fermentation substrate, and relative usage is larger, the shape generally added using batch
Formula is added in fermentation system, therefore the sterilizing of liquid alkane generally requires carry out bulk sterile.For example produced in long-chain biatomic acid
In, specially prepare liquid alkane storage tank, the alkane required for whole fermentation period has been prestored, and carry out sterilization treatment.
Temperature is mainly risen to 121 DEG C, and constant temperature pressurize 30min by the sterilizing of liquid alkane using steam-heated mode.
In disinfecting action, warming temperature generally is carried out by the way of chuck heating, but chuck programming rate is slower, therefore typically
In temperature-rise period heating heating can be carried out in the way of logical steam.Such operation can form larger in the prometaphase of sterilizing
Evaporation capacity, so as to carry substantial amounts of alkane secretly in steam, statistics show this alkane loss account for alkane usage amount 10%~
20%.Meanwhile, the mode that bottom leads to steam makes steam directly be contacted with alkane, therefore can form substantial amounts of condensed water and mix in alkane
In hydrocarbon, emulsion layer is formed, water environment is provided for the intrusion of miscellaneous bacteria, improves microbiological contamination risk.Under existence conditions, no matter to this
What kind of optimization sterilization method carries out, and can all cause alkane to lose and formed aqueous phase emulsion layer, so as to cause certain to fermentation
Influence.
The content of the invention
In view of the shortcomings of the prior art, the present invention provides a kind of sterilizing methods of liquid alkane.This method is by improving liquid
The sterilization process of state alkane, reduces the evaporation loss of alkane and the formation of emulsion layer, reduces sterilising temp, shorten sterilizing
Time, improve sterilizing efficiency.
The sterilizing methods of the liquid alkane of the present invention, including herein below：For the liquid alkane of bioconversion, through microwave
Demulsification is handled, alkane and water stratification, adds bacteriostatic agent, carries out microwave sterilization processing, the liquid alkane sterilized.
In the inventive method, described liquid alkane is the alkane containing 5~16 carbon atoms that can be used in biofermentation
Hydrocarbon, is such as used for 12 carbon n-alkanes, 14 carbon n-alkanes that long-chain biatomic acid ferments.
In the inventive method, the condition of microwave demulsifying processing is：Microwave frequency is 300~3000MHz, Microwave Power Density
For 100W/L~10kW/L, treatment temperature is 50 ~ 90 DEG C, and processing time is 10 ~ 30 minutes.
In the inventive method, microwave demulsifying processing is preferred to use the mode of intermittent-heating, is specially：It is warming up to 70 first~
90 DEG C, then stop microwave, naturally cool to 50~60 DEG C, 2~4 demulsification operations are carried out in this way.
In the inventive method, one kind in for monosaccharide and disaccharide and acylate, inorganic salts of described bacteriostatic agent or
Several, wherein monose is glucose, fructose, galactolipin, ribose etc., preferably glucose；Disaccharide is sucrose, lactose, maltose etc.,
Preferably sucrose；Acylate is citrate, acetate, amino-acid salt etc., preferably sodium acetate；Inorganic salts are sodium chloride, chlorination
Potassium etc., preferably sodium chloride；The addition of bacteriostatic agent is 1 with the mass ratio of liquid alkane reclaimed water：0.5~1：10, preferably 1：2~1：5.
In the inventive method, the condition of microwave sterilization processing is：Microwave frequency is 300~3000MHz, Microwave Power Density
For 20~100W/L, processing time is 5~10 minutes.
Compared with prior art, the inventive method has the following advantages that：
（1）Sterling is not easy microbiological contamination situation occur, but often carries certain moisture secretly in the liquid alkane of reality, from
And be that bacteria breed brings certain possibility, therefore alkane sterilizing generally be directed to this part water entrainment, present invention side
Method introduces two kinds of control devices of microwave and bacteriostatic agent, and sterilizing is targeted, while being cooperated between microwave and bacteriostatic agent, goes out
Bacterium effect is protruded, the problem of solving alkane loss present in conventional steam sterilizing and high microbiological contamination risk.
（2）Alkane density ratio water is small, therefore in alkane system, stratification state of the alkane in upper water under, water tool occurs
Polarized, there is good wave-absorbing property, by way of microwave is heated, and the temperature of water entrainment can be instantaneously improved, in microwave
Under effect, the effect that can be sterilized to aquatic products heat, simultaneously as the apolar character of alkane, does not have wave-absorbing property, is conducive to
The stripping of combination water in alkane.
（3）The bactericidal action of microwave mainly by making protein denaturation, changing the modes such as permeability of cell membrane, influences thalline
Eubolism, so as to play bactericidal action.But bacterium itself has autonomous adjusting function as life entity, copes with external
Stimulate, form stress reaction.The basis of this regulating and controlling effect is water environment.This method is by adding monosaccharide and disaccharide, organic acid
The additive such as salt and inorganic salts, forms local hypertonic environment in alkane system, so as to can make the moisture in somatic cells
It is a large amount of to be lost in, make thalline stress mechanism be suppressed, so as to greatly improve the bactericidal effect of microwave.Meanwhile, these are highly concentrated
The nutriment of degree can be entered in fermentation system as alkane stream adds, so as to promoting fermentation strain metabolism, improve fermentation water
It is flat, with certain gain effect.
Embodiment
The detailed process and effect of the inventive method are illustrated with reference to embodiment, but are not limited to following implementation
Example.
Candida tropicalis is selected in the present embodiment（Candida tropicalis）Mutant strain PF-UV-56 is used as fermentation
Bacterial strain carries out the experiment of long chain alkane fermenting and producing long-chain biatomic acid, and the mutant strain is deposited in Chinese microorganism strain preservation management committee
Member's meeting common micro-organisms center, deposit number is CGMCC NO.0356.Using karl Fischer drimeter in alkane system
Water content measure, so that it is determined that each state modulator that sterilizes.
Embodiment 1
（1）By the carbon n-alkanes of 10L 12, water content 2%（V:V）, add in 15L feed supplement storage tanks, microwave demulsifying processing,
Microwave frequency 2450MHz, microwave power is 5KW, and microwave is heated to 90 DEG C, is then naturally cooling to 50 DEG C, repeatedly heat up-
Temperature-fall period is twice.
（2）To step（1）In alkane in add glucose 200g, carry out microwave sterilization, microwave frequency 896MHz, microwave
Power is 200W, is handled 10 minutes, and the alkane after sterilizing is designated as A1, is cooled standby.
Embodiment 2
（1）By the carbon n-alkanes of 10L 12, water content 3.3%（V:V）, add in 15L feed supplement storage tanks, at microwave demulsifying
Reason, microwave frequency 896MHz, microwave power is 1KW, and microwave is heated to 80 DEG C, is then naturally cooling to 60 DEG C, repeats to rise
Temperature-temperature-fall period 4 times.
（2）To step（1）In alkane in add sucrose 80g, carry out microwave sterilization, microwave frequency 500MHz, microwave work(
Rate is 400W, is handled 10 minutes, and the alkane after sterilizing is designated as A2, is cooled standby.
Embodiment 3
（1）By the carbon n-alkanes of 10L 12, water content 5.2%（V:V）, add in 15L feed supplement storage tanks, at microwave demulsifying
Reason, microwave frequency 500MHz, microwave power is 1KW, and microwave is heated to 90 DEG C, is then naturally cooling to 50 DEG C.
（2）To step（1）In alkane in add sodium acetate 66g, carry out microwave sterilization, microwave frequency 400MHz, microwave
Power is 400W, is handled 10 minutes, and the alkane after sterilizing is designated as A3, is cooled standby.
Comparative example 1
By the carbon n-alkanes of 10L 12, water content 2%（V:V）, add in 15L feed supplement storage tanks.Chuck is heated to 90 DEG C, with
Tank bottom is passed through 0.4MPa steam afterwards, is warming up to 121 DEG C, maintains temperature after 30 minutes, naturally cools to 37 DEG C.Alkane after sterilizing
It is designated as A4.
Comparative example 2
By the carbon n-alkanes of 10L 12, water content 3.3%（V:V）, add in 15L feed supplement storage tanks, chuck is heated to 90 DEG C,
Subsequent tank bottom is passed through 0.4MPa steam, is warming up to 121 DEG C, maintains temperature after 30 minutes, naturally cools to 37 DEG C.Add sucrose
80g, stirs.This alkane is labeled as A5.
Comparative example 3
By the carbon n-alkanes of 10L 12, water content 5.2%（V:V）, add in 15L feed supplement storage tanks, microwave demulsifying processing is micro-
Wave frequency rate 500MHz, microwave power is 1KW, and microwave is heated to 90 DEG C, is then naturally cooling to 50 DEG C, repeats heating-drop
Twice, the alkane after sterilizing is labeled as A6 to warm process.
Embodiment 4
Using candida tropicalis as fermentation strain, respectively using 12 carbon n-alkane A1 ~ A6 after sterilizing as substrate, use
Batch is added, gradient regulation pH mode carries out aerobic fermentation, until reaction terminates during 144h.Final fermentation results are shown in Table 1.
The result of the SL-AH fermentation test of table 1.
| Water content before sterilizing（%） | Water content after sterilizing（%） | Batch fermentation | Averagely produce acid concentration（g/L） | The preservation cycle（My god） | |
| A1 | 2 | 2 | 4 | 150.1 | 90 |
| A2 | 3.3 | 3.3 | 4 | 148.6 | 90 |
| A3 | 5 | 5 | 4 | 142.1 | 90 |
| A4 | 2 | 6.5 | 3 | 127.5 | 78 |
| A5 | 3.3 | 7.6 | 3 | 134.4 | 72 |
| A6 | 5 | 5 | 4 | 135.9 | 83 |
12 carbon n-alkane liquid storages of sterilization treatment are carried out in the process of the present invention（A1、A2）, water content will after it sterilizes
Significantly lower than conventional wet heat disinfection（A4、A5）, while the loss of alkane is less, batch fermentation is higher than control group；Due to this
The additives such as monosaccharide and disaccharide, acylate and inorganic salts are with the addition of in inventive method, the nutriment of alkane system is optimized
Constitute, its fermentation level is significantly better than control group, and coordinates the sterilizing effect of microwave, and substantially, the preservation cycle is longer for sterilization effect.
Claims (8)
1. a kind of sterilizing methods of liquid alkane, it is characterised in that including herein below：For the liquid alkane of bioconversion, warp
Microwave demulsifying processing, alkane and water stratification, add bacteriostatic agent, carry out microwave sterilization processing, the liquid alkane sterilized；It is described
One or more of the bacteriostatic agent in for monosaccharide and disaccharide and acylate, inorganic salts.
2. in accordance with the method for claim 1, it is characterised in that：Described liquid alkane is that can be used in containing for biofermentation
There is the alkane of 5~16 carbon atoms.
3. in accordance with the method for claim 1, it is characterised in that：Microwave demulsifying processing condition be：Microwave frequency be 300~
3000MHz, Microwave Power Density is 100W/L~10kW/L, and treatment temperature is 50 ~ 90 DEG C, and processing time is 10 ~ 30 minutes.
4. according to the method described in claim 1 or 3, it is characterised in that：Microwave demulsifying processing is by the way of intermittent-heating, tool
Body is：70~90 DEG C are warming up to first, then stops microwave, 50~60 DEG C are naturally cooled to, and 2~4 times are carried out in this way and is broken
Breast operation.
5. in accordance with the method for claim 1, it is characterised in that：Described monose is glucose, fructose, galactolipin or ribose
In one or more；Disaccharide is the one or more in sucrose, lactose or maltose；Acylate is citrate, acetic acid
One or more in salt or amino-acid salt；Inorganic salts are sodium chloride or potassium chloride.
6. in accordance with the method for claim 1, it is characterised in that：The addition of bacteriostatic agent and the mass ratio of liquid alkane reclaimed water
For 1：0.5~1：10.
7. in accordance with the method for claim 1, it is characterised in that：The addition of bacteriostatic agent and the mass ratio of liquid alkane reclaimed water
For 1：2~1：5.
8. in accordance with the method for claim 1, it is characterised in that：Microwave sterilization processing condition be：Microwave frequency be 300~
3000MHz, Microwave Power Density is 20~100W/L, and processing time is 5~10 minutes.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1821075A (en) * | 2006-03-01 | 2006-08-23 | 汪平华 | Method for producing organic clay |
| CN102648168A (en) * | 2007-06-12 | 2012-08-22 | 英国石油国际有限公司 | Process for converting ethane into liquid alkane mixtures |
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