CN108117995A - A kind of preparation method of sweet potato bacterium culture medium - Google Patents

Abstract

The present invention relates to a kind of preparation method of sweet potato bacterium culture medium, agricultural technology field is by following step progress：First, the preparation of raw material：New fresh sweet potatoes, agar, water；2nd, sweet potato spontaneous fermentation is decomposed：27 DEG C~30 DEG C of fermentation temperature, air humidity 20g~25g/m³, itself 13~15 days resolving times of standing for fermentation generation carbohydrate；3rd, sweet potato juice makes：Sweet potato is thinly sliced, boiling is added to boil 7~8 minutes, sweet potato juice is obtained with filtered through gauze after cooling；3rd, agar is added in sweet potato juice, rise temperature to 80 DEG C~100 DEG C dissolvings becomes mixed liquor；4th, 50 DEG C~70 DEG C time-divisions of mixed liquor are instilled in ten test tubes；5th, medium sterilization：A bundle of test tube for having dispensed mixed liquor is put into together, high-temperature sterilization is carried out in sterilization tank；6th, the formation of medium slant：A bundle of test tube after having sterilized takes out and plane places after being cooled to 15 DEG C~25 DEG C into 30-degree angle and is frozen into slant medium.

Description

A kind of preparation method of sweet potato bacterium culture medium

Technical field

The present invention relates to a kind of production methods of sweet potato bacterium culture medium, belong to agricultural technology field.

Background technology

Strain is the microorganism as living-cell catalyst for fermentation process, including bacterium, actinomyces, saccharomycete and mould Four major class of bacterium.From the substantial amounts of microorganism of nature, therefrom through separating and filter out useful strain, then improved, stored It is ready to use in production.Strain refers to the propagating materials of edible mushroom, industrial bacterium, agricultural bacterium mycelium and its growth substrate composition.Strain It is divided into parent species (level-one kind), original seed (two level kind) and cultigen (three-level kind) three-level.

In March, 2006 The Ministry of Agriculture of the People's Republic of China, MOA's issue《Edible fungus species management method》Regulation：" Ministry of Agriculture master The strain work of the pipe whole nation.Agricultural (edible mushroom, similarly hereinafter) administrative responsibile institution of local people's governments at or above the county level is responsible for this administrative area Bacterial Strains Managing work in domain." " local people's governments at or above the county level administrative department in charge of agriculture should strengthen edible mushroom germplasm money Source protection and fine-variety breeding, production, update, popularization encourage selection and breeding, production, operation to be combined." industrial fermentation it is useful Strain, screening step include strain separating, primary dcreening operation and secondary screening, select the useful strain with certain ability.

It is the collection sample containing bacterium from soil or saprophyte first, after being diluted with sterile water, is coated on and is equipped with suitable for thin It on the agar culture plate of bacterium, actinomyces or fungus growth, and is inverted in insulating box, cultivates certain time, plate On many single bacterium colonies (colony of single microorganism) for growing be various purebred bacterial strains after separating respectively, it is referred to as purebred； On culture transferring to test tube slant culture medium, it is spare to put 4 DEG C of refrigerators.

According to different screening purposes, using different screening models.As the conventional method for screening antibiotics generated bacterium is Will be separating obtained purebred in soil, after being cultivated on the plate containing agar medium, with card punch by fungus block move to containing It on the agar medium of test organisms, cultivates take out after a certain period of time at a suitable temperature, such as have around fungus block transparent Inhibition zone then shows that this strain has the ability for generating the antibacterial material for inhibiting test organisms growth.Gained strain passes through shaking flask liquid Body ferments, and measures the content of antibiotic in zymotic fluid or mycelium, selects the high strain of production capacity.On the other hand to being extracted Antibiotic carry out structural analysis, pharmacological testing and clinical test etc., be really responder, you can as production strain.For another example sieve The method for selecting lipase production strain is that separating obtained mould is coated on the agar medium containing tallow, constant temperature incubation Certain time, as transparent circle occurs in periphery of bacterial colonies, then the bacterium has lipolytic ability, further makees shake flask fermentation measure Enzyme activity selects the starting strain that the high person of yield makees production strain.

Strain improvement that is, using the method for genetic breeding, makes wild-type strain (from nature separation screening obtained by Bacterial strain) gene DNA undergo mutation, recombinate, so as to therefrom select yield high, quality of finished product good or special with new culture Property for example resistance to Product inhibiton, can utilize cheap raw material and with production new varieties ability excellent species.The method of use lures Become breeding, crossbreeding, cell-fusion techniques and recombinant DNA technology.Mutation breeding be using mutagen such as ultraviolet light, cobalt- 60th, the monospore suspension of the physically or chemically mutagens process for producing bacterial strain such as the aziridine type, to obtain Mutation induction strain.With The screening of mutant strain is carried out afterwards, therefrom screens superior strain.Since in random mutagenized populations, beneficial mutation proportion is very Low, to obtain high productive mutant must largely be screened.It in recent years, can basis with the development of fermentating metabolism control research Reflecting point in biosynthesis pathway, and pass through their change to improve yield or other characteristics, such as the anti-product feedback of selection and breeding The mutant strain of inhibition, the mutant strain for increasing cell permeability and auxotrophic mutant strain etc..This " rational screening method " is extensive Selection and breeding applied to Amino Acid-producing Bacteria.

Prepared by strain, also referred to as prepared by seed, refers to strain under certain condition, has a fixed number by expanding to cultivate to become The preparation process of the pure production strain of amount and quality.With make further to expand in access fermentation tank biomass and synthetic product it With.Seed preparation includes spore and prepares and mycelium preparation.Mycelial preparation generally takes Tube propagation base to carry out fructification Inoculation, the production strain being stored in sandy soil pipe or cryovial is a little with sterile formality picking, accesses agar slant culture-medium On, 5~7 days (or long period) is cultivated under 25 DEG C (or higher temperature).Gained spore also needs further to use large surface area Solid medium to obtain more spores, prepared for der Pilz spore, majority using rice, millet etc natural cultures Base.

The content of the invention

In order to prepare a kind of culture medium for being more suitable for bacterium multiple eating strain, solve some strains and be difficult to asking for artificial cultivation Topic, a kind of production method of sweet potato bacterium culture medium of my invention, this method do not have to add in sugar, and sweet potato fermentation is allowed to generate Sugar is further carried out preparing, and culture medium obtained more can make mycelia survive and breed faster.

A kind of preparation method of sweet potato bacterium culture medium solve the problems, such as to adopt the technical scheme that by following step into Row：First, the preparation of culture medium raw material：New one 150 grams of fresh sweet potatoes, 20 grams of agar, 1000 grams of water；2nd, sweet potato spontaneous fermentation point Solution：Fresh sweet potato is cleaned, is put into fermentation Porcelain Jar, keeps 27 DEG C~30 DEG C of temperature, air humidity 20g~25g/m³, from Body 13~15 days resolving times of standing for fermentation generate carbohydrate；3rd, sweet potato juice makes：Sweet potato from jar fermenter is taken out, is removed the peel, is cut Into the thin slice of 0.3cm~0.4cm thickness, 1000 grams of water is added in, boils 7~8 minutes, is smashed to pieces when boiling, with three layers after Temperature fall Gauze encirclement pinches press filtration and obtains sweet potato juice；3rd, agar is added in sweet potato juice, rise temperature is to 80 DEG C~100 DEG C, Bian Jia Hot side stirs evenly, until fully dissolving becomes mixed liquor；4th, take ten test tubes cleaned are parallel to be bundled, be disposed vertically, It is drawn, point instilled in ten test tubes using dropper during 50 DEG C~70 DEG C of mixed liquor, every test tube drips loading amount 1/5, every after the completion of drop Branch test tube uses cotton ball stoppered test tube mouth respectively, and cotton ball is unable to contact liq；5th, medium sterilization：Mixed liquor will have been dispensed A bundle of test tube put into high-temperature sterilization is carried out in sterilization tank together, temperature is slowly raised, when reaching 90 DEG C~100 DEG C of sterilising temp 60 DEG C are naturally cooling to after being kept for 20~30 minutes；6th, the formation of medium slant：A bundle of test tube after having sterilized takes out, Temperature is not less than 60 DEG C during taking-up, while hot places this bundle test tube bottom surface and plane into 30-degree angle, makes in test tube liquid level into oblique Face, standing are frozen into slant medium after being cooled to 15 DEG C~25 DEG C.

A kind of advantageous effect of the preparation method of sweet potato bacterium culture medium is：A kind of preparation method of sweet potato bacterium culture medium It is by the way that sweet potato is stood spontaneous fermentation decomposition generation monose and polysaccharide, sweet potato is ripe using sweet potato and agar, water as raw material It mixes, be in charge of and sterilize and the shaping of medium slant with agar after change, slant medium is made；A kind of sweet potato strain training The preparation method for supporting base carries out body fermentation using the moisture of itself in new fresh sweet potatoes and starch, and generation can more be suitble to growth Substance, solve the problems, such as that some rare strains are difficult to manually cultivate.

Specific embodiment

Embodiment：A kind of preparation method of sweet potato bacterium culture medium is carried out by following step：First, the standard of culture medium raw material It is standby：New one 150 grams of fresh sweet potatoes, 20 grams of agar, 1000 grams of water；2nd, sweet potato spontaneous fermentation is decomposed：Fresh sweet potato is cleaned, It is put into fermentation Porcelain Jar, keeps 27 DEG C~30 DEG C of temperature, air humidity 20g~25g/m³, itself standing for fermentation resolving time 13 Generate carbohydrate within~15 days；3rd, sweet potato juice makes：Sweet potato from jar fermenter is taken out, is removed the peel, is cut into the thin of 0.3cm~0.4cm thickness Piece adds in 1000 grams of water, boils 7~8 minutes, smashed to pieces when boiling, surrounded after Temperature fall with three layers of gauze pinch press filtration obtain it is red Potato juice；3rd, agar is added in sweet potato juice, rise temperature heats while stirring uniformly to 80 DEG C~100 DEG C, until filling Dissolving is divided to become mixed liquor；4th, take ten clean test tubes are parallel to be bundled, be disposed vertically, in 50 DEG C~70 DEG C of mixed liquor It is drawn using dropper, in point ten test tubes of instillation, every test tube drips loading amount 1/5, and every test tube uses cotton ball respectively after the completion of drop Stoppered test tube mouth, cotton ball are unable to contact liq；5th, medium sterilization：A bundle of test tube for having dispensed mixed liquor is put into together High-temperature sterilization is carried out in sterilization tank, slowly raises temperature, after being kept for 20~30 minutes when reaching 90 DEG C~100 DEG C of sterilising temp from So it is cooled to 60 DEG C；6th, the formation of medium slant：A bundle of test tube after having sterilized takes out, and temperature is not less than 60 during taking-up DEG C, this bundle test tube bottom surface and plane into 30-degree angle is placed while hot, makes in test tube liquid level into inclined-plane, standing be cooled to 15 DEG C~ Slant medium is frozen into after 25 DEG C.

Claims (1)

1. a kind of preparation method of sweet potato bacterium culture medium, it is characterized in that being carried out by following step：First, the standard of culture medium raw material It is standby：New one 150 grams of fresh sweet potatoes, 20 grams of agar, 1000 grams of water；2nd, sweet potato spontaneous fermentation is decomposed：Fresh sweet potato is cleaned, It is put into fermentation Porcelain Jar, keeps 27 DEG C~30 DEG C of temperature, air humidity 20g~25g/m³, itself standing for fermentation resolving time 13 Generate carbohydrate within~15 days；3rd, sweet potato juice makes：Sweet potato from jar fermenter is taken out, is removed the peel, is cut into the thin of 0.3cm~0.4cm thickness Piece adds in 1000 grams of water, boils 7~8 minutes, smashed to pieces when boiling, surrounded after Temperature fall with three layers of gauze pinch press filtration obtain it is red Potato juice；3rd, agar is added in sweet potato juice, rise temperature heats while stirring uniformly to 80 DEG C~100 DEG C, until filling Dissolving is divided to become mixed liquor；4th, take ten clean test tubes are parallel to be bundled, be disposed vertically, in 50 DEG C~70 DEG C of mixed liquor It is drawn using dropper, in point ten test tubes of instillation, every test tube drips loading amount 1/5, and every test tube uses cotton ball respectively after the completion of drop Stoppered test tube mouth, cotton ball are unable to contact liq；5th, medium sterilization：A bundle of test tube for having dispensed mixed liquor is put into together High-temperature sterilization is carried out in sterilization tank, slowly raises temperature, after being kept for 20~30 minutes when reaching 90 DEG C~100 DEG C of sterilising temp from So it is cooled to 60 DEG C；6th, the formation of medium slant：A bundle of test tube after having sterilized takes out, and temperature is not less than 60 during taking-up DEG C, this bundle test tube bottom surface and plane into 30-degree angle is placed while hot, makes in test tube liquid level into inclined-plane, standing be cooled to 15 DEG C~ Slant medium is frozen into after 25 DEG C.