CN1641016A

CN1641016A - Composite culture medium for producing phosphorus-potassium dissolving bacteria and its liquid deep fermentation producing method

Info

Publication number: CN1641016A
Application number: CN 200410002185
Authority: CN
Inventors: 郭伟光
Original assignee: Individual
Current assignee: Individual
Priority date: 2004-01-14
Filing date: 2004-01-14
Publication date: 2005-07-20

Abstract

The present invention is composite culture medium for producing phosphate and potash decomposing bacteria and its deep liquid fermentation producing process. The composite culture medium comprises corn saccharifying liquid, red sugar, honey, silkworm cocoon powder, lecithin, potash feldspar, inorganic salts and composite vitamins. In the fermentation process, sectional control is adopted to control the ventilation amount, stirring speed and culture temperature for promoting phosphate and potash decomposing bacteria growth, strengthening the sync growth of spore and avoiding microbial contamination and. The present invention may be used in producing bacterial fertilizer products with great amount of live phosphate and potash decomposing bacteria, high spore growing rate and long preservation period.

Description

Produce complex medium and the liquid submerged fermentation production method thereof of P and K decomposing bacterium

The present invention relates to a kind of complex medium and liquid submerged fermentation production method thereof of producing the P and K decomposing bacterium, belong to the microbial fertilizer production field.

Microbial fertilizer quality quality key is two aspects, the one, and the quality of bacterial classification comprises the nitrogen fixing capacity, phosphorus decomposing ability, ability of dissolving potassium of bacterial classification, to the adaptive faculty of severe environment etc.; The 2nd, amount of viable bacteria comprises quantitative proportion between the bacterial classification, the whether variation of number of viable etc. between antagonism, preservation period mutually, and substratum and production method that these two aspects are adopted during with bacterial manure production are closely-related.In the present disclosed microbial fertilizer making method, generally adopt all kinds of bacterial classifications to cultivate the decomposite mode of production respectively, this method complex operation causes living contaminants easily, is difficult to ensure the quality of product; The mode of the employing multiple bacteria compound fermentation that also has, but because adopt general substratum, be difficult to fully ensure all kinds of bacterial classifications nutritional needs separately, therefore though total count has reached standard in the product that comes out of fermentative production, but all kinds of bacterial classification quantity differ, ratio is inharmonious, can not embody the physiological action of all kinds of active bacterias fully; What also have class employing in addition is the mode of solid fermentation, and the product viable count that this method is produced is generally not high, has only reached the quantity of expection in other words in theory.

The object of the present invention is to provide the complex medium of a kind of suitable production P and K decomposing bacterium.

Another object of the present invention is to provide a kind of liquid submerged fermentation production method of cultivating the P and K decomposing bacterium.

Complex medium of the present invention is made up of crude substance, inorganic salt and vitamin complex, is particularly suitable for the mixed culture of phosphorus bacteria and potassium bacterium.The major ingredient of substratum is:

Corn saccharification liquid 15% brown sugar 2% honey 0.5%

Silk cocoon powder 0.5% Yelkin TTS, 0.02% potassium felspar sand or mica 0.1%

KH ₂PO ₄ 0.2％ CaCO ₃ 0.25％ MgSO ₄ 0.02％

FeSO ₄7H ₂O 10PPM MnSO ₄H ₂O 5PPM vitamin complex 10PPM

Bacterial classification of the present invention is provided by Chinese common micro-organisms DSMZ, one is phosphate solubilizing bacteria, numbering AS 1.220, one is bacillus megaterium, numbering AS 1.217, Gram-positive produces oval gemma, can observe tangible molten phosphorus circle containing on the agar plate of Yelkin TTS to cultivate; Other one is colloid bacillus cereus, numbering AS 1.231, and Gram-negative has pod membrane, and oval gemma is arranged.These three bacterial classifications are equal well-grown on nitrogen-free agar, all has certain nitrogen fixation.

Zymotechnique of the present invention adopts sectionalized control method, promotes thalli growth by ventilation, stirring velocity and culture temperature in the control fermenting process, strengthens the synchronous production rate of gemma.Earlier fermentation and later stage ventilation are little, and the ferment middle ventilation is big, and the fermentation later stage is improved culture temperature.

Fermentative production early stage, bacterial classification carries out oneself adjustment under new nutritional condition, be mainly reflected in the variation of thalline volume size on the macroscopic view, and the variation of thalline quantity is not obvious, and thalli growth is little to the demand of oxygen, so can reduce ventilation during this period of time; Ferment middle, thalli growth is vigorous, and quantity is with geometric growth, and thalline at this moment is big to the demand of oxygen, needs to strengthen ventilation and carries out oxygen supply, can improve the rotating speed of stirring simultaneously, to improve dissolved oxygen level and to accelerate flowing of nutritive substance.In the fermentation later stage, the bacterium number tends towards stability substantially, and gemma begins a large amount of formation, and can reduce ventilation this time gradually, and improves culture temperature to 45～50 ℃ simultaneously, helps the synchronous formation of thalline endospore like this.Facts have proved that phosphorus bacteria that comes out with this segmentation control fermentative production and the gemma production rate of potassium bacterium be more than 98%, thereby make the validity period of bacterial manure prolong greatly.

Complex medium of the present invention and liquid submerged fermentation production method be what sum up in bacterial manure production practice for many years, passed through repeatedly facts have proved and be better than general common production substratum and production method that its characteristics are:

1. substratum comprehensive nutrition, collocation rationally, the fast growth of bacterial classification just can be finished fermentation in general about 28～32 hours, the viable count in every milliliter of fermented liquid is more than 2,500,000,000;

2. phosphorus bacteria and potassium bacterium all can stable growths in the fermented liquid process, and eurythmy was inoculated than 1: 2 by quantity as phosphorus bacteria and potassium bacterium, and the bacterial fertilizer products bacterium number of then producing is stabilized in about 1: 1.5 than generally;

3. fermentation is carried out in closed environment, and is time saving and energy saving, stopped living contaminants, and the quality of bacterial manure has obtained guarantee;

4. realized the synchronization that gemma generates, gemma production rate height, thus make bacterial fertilizer products that longer preservation period be arranged.

The present invention is further illustrated with embodiment below:

1. prepare corn saccharification liquid

Take by weighing and be ground into 150 kilograms of 80 purpose Semen Maydis powder, add in 300 kg of water, be warmed up to 65 ℃, add saccharifying enzyme, insulation saccharification 3～5 hours, dripping iodine liquid to the saccharification liquid, not show blueness be terminal point.The complete after-filtration of saccharification gets saccharification liquid.

2. fermentative production

The corn saccharification liquid that makes is changed in the fermentor tank, add 20 kilograms in brown sugar, 5 kilograms of honey, 5 kilograms in silk cocoon powder, 0.2 kilogram in Yelkin TTS, 1 kilogram of potassium felspar sand fine powder, KH ₂PO ₄2 kilograms, CaCO ₃2.5 kilogram, MgSO ₄0.2 kilogram, FeSO ₄7H ₂O 10 grams, MnSO ₄H ₂O 5 grams, vitamin complex 10 grams are warming up to 110 ℃ of autoclavings, are cooled to 30 ℃ again, insert bacterial classification.Inoculum size is 5%～8%, and phosphorus bacteria and potassium bacterial number ratio control are about 1: 2 in the bacterial classification.Preceding 6～8 hours ventilations of fermenting are controlled at 1: 0.3～0.35; After 6～8 hours, thalline progressively enters logarithmic phase, in great demand to oxygen and nutritive substance, need to strengthen ventilation during this period of time, to quicken flowing of dissolved oxygen and nutritive substance, the ventilation size can be decided according to fermentative production, generally between 1: 0.5～0.65, also can change according to the production practical situation.In the later stage of fermentative production, gemma begins to form, and at this moment will reduce ventilation, helps the formation of gemma like this.Culture temperature is brought up to 45 ℃, forms synchronously to impel gemma.The general cultivation about 30 hours, leavened prod is qualified through microscopy, fermentation ends.

With the leavened prod that is up to the standards through sterile filling, just obtained liquid bacterial fertilizer; As fermented liquid is adsorbed with the aseptic peat composed of rotten mosses, just obtained solid bacterial manure.

Claims

1. a complex medium and liquid submerged fermentation production method thereof of producing the P and K decomposing bacterium is characterized in that the major ingredient of complex medium is:

Corn saccharification liquid 15% brown sugar 2% honey 0.5%

Silk cocoon powder 0.5% Yelkin TTS, 0.02% potassium felspar sand or mica 0.1%

KH ₂PO ₄ 0.2％ CaCO ₃ 0.25％ MgSO ₄ 0.02％

FeSO ₄7H ₂O 10PPM MnSO ₄H ₂O 5PPM vitamin complex 10PPM

2. liquid submerged fermentation production method according to claim 1, it is characterized in that adopting sectionalized control method, promote thalli growth by ventilation, stirring velocity and culture temperature in the control fermenting process, strengthen the synchronous production rate of gemma, earlier fermentation and later stage ventilation are little, the ferment middle ventilation is big, and the fermentation later stage is improved culture temperature.

3. P and K decomposing bacterium according to claim 1 is characterized in that bacterial classification of the present invention is provided by Chinese common micro-organisms DSMZ, and one is phosphate solubilizing bacteria, and numbering is respectively AS 1.220; One is bacillus megaterium, numbering AS 1.217; Other one is colloid bacillus cereus, numbering AS 1.231.