Fermenti. Characteristics of soil enzymes. Yak work in organizmі
Understanding fermentation
Enzymes (enzymes) I call it the rosette or tied with the membranes of the bricks, over the catalytic activity. (Крім білків catalytic activity organisms can show actions of RNA (ribozymes) and antibodies (abzymes), however, the stench in a thousand times less effective, less ferment.) Smell stench about persh_ dzherelakh enzymes. Biohimii, yaka vivchak ferment, be called enzymology... On the diagrams і in the reactions of the enzyme molecules mean - E... Recovini, a re-enactment of what catalyzes ferments, is called substrates (S). products enzyme reactions mean - R... In addition, ferments can be prepared in a homogeneous glass in the same ways, as well as in іnshі bottles. For enzymes, they are characterized by physical and chemical power, power to the little ones.
Specification of enzymes from inorganic catalysts:
a) accelerate the reaction in a meaningfully effective way;
b) over a high specificity of the work;
c) subject to regulation in physiological minds;
d) dіyut in soft minds.
budova ferments
Enzymes can be used as simple or folding (kon'yugovany) bottles, before the warehouse of which can include lipid, carbohydrates, metals, nitrogenous bases, old vitamins. In organism, enzymes can function as a function of a plant, as well as in a non-essential complex, or it enters the storehouse of biological membranes.
Due to the specificity of enzymes active center. Active center - a very unique combination of proximity in the space of amino acid surplus, which will prevent:
a) recognition of the molecule to the substrate,
b) linking the substrate with the enzyme,
c) the catalytic conversion (in the case of a folding enzyme in the act of catalysis, it also takes care of the fate of the coenzyme, which enters the active center before the warehouse).
The active center is winnowed at the moment when the blocks turn off and accept its native (active) conformation. The structure of the active center can change during interaction with the substrate. Behind the figurative vislov of D. Koshland, the substrate goes to the active center, yak hand to the mittens.
One molecule of the enzyme, especially if it is stored in a decimal subunit, can be attached to more than one active center.
In the active center є dvі dilyanka. The first dilyanka is prompted for recognition and connection to the substrate. Vin is called a substratum-sounding dilyanka or an anchored maidan. Another dilyanka is called catalytic, its warehouse includes amino acid surpluses, which take part in the act of catalysis.
Enzymes are proteins that are strongly developed for molecular weight and folding. Applying the enzyme with a small molecule є ribonuclease, it can be stored in one subunit with a molecular weight of 13700 Da. (Ribonuclease has an amino acid endurance. In 1969, ribonuclease was synthesized in B.Merrifild's laboratory in New York.) Bagato enzymes are stored in one sub-unit, for example, lactate dehydrogenase is two subtypes. Until the present hour, there are a number of multi-enzyme complexes, which are stored in dozens of different sub-units and decile types of coenzymes. For example, the pyruvate dehydrogenase complex consists of 60 subunits of three types and five types of cofactors. The molecular weight of such a complex becomes 2.3 * 10 6 - 10 * 10 6 Da in the presence of dzherel to the enzyme. The enzyme molecule can be smaller than the substrate molecule. For example: molecules of enzymes amylase and ribonuclease are less, less molecules of substrates - starch and RNA.
The bilkov part of folding enzymes is catalytically inactive and is called apofenzyme... The connection of the apoenzyme with a non-alcoholic component is to be produced until the catalyst is approved for the catalytically active enzyme (holoenzyme):
Bagato fermentations in their own warehouses for metal, which may be viconuvati development functions:
a) take part in the involved substrate and the process of its catalytic transformation;
b) take the attached coenzyme to the enzyme molecule;
c) stabilize the tretin structure of the enzyme (for example, Ca 2+ in amylase);
d) vibrate with the substrate, fix the true substrate, for which enzyme.
A lot of coenzymes є obsolete vitamins, the failure of the exchange of speech in case of vitamic deficiencies is explained by a decrease in the activity of singing enzymes.
Deyaki ferments order with the active center to revenge allosteric (regulatory) center - dilyanka of a bilkovo globule, a pose to the active center, can sound the words, and regulate the enzymatic activity. The words of speech are called allosteric effectors (allosteric activators or ingibitors)... As a result of the linking of the effector to the allosteric center, a change in the structure of the bottle is observed, which leads to a change in the spacious growth of amino acid surpluses in the active center, as a result, to a change in the enzymatic activity.
Enzymes that are created in one organism and catalyze one or the same cheeky reaction Alle with the original structure of the protein, they are called isoenzymes. Ferments are produced one kind one after such physical and chemical authorities, Yak molecular weight, thermal stability, Substrate specificity, electrophoretic looseness. The nature of the appearance of isoenzymes is versatile, but more often for all it is amazed by the ideas in the structures of genes, which code isoenzymes or subunits. For example, the enzyme lactate dehydrogenase (LDH), which catalyzes the reverse reaction of oxidized lactate to pyruvate, may be subunits of two types M and H, the combination of these subunits is the basis of the formation of five Lizoenzymes. For the diagnosis of heart disease and liver disease, it is necessary to monitor the LDH isozyme spectrum in the blood, LDH 1 and LDH 2 actives in the heart and blood cells, and LDH 4 and LDH m 5 - as early as in the summer.
Fig. 1 Budova LDH isozymes.
Vimіryuvannya enzymatic activity
The value of the activity of enzymes is going to go by the way. The frequency of enzymatic reactions is varied by decreasing the concentration of the substrate or increasing the concentration of the product per hour:
v = -ΔC S / Δτ, v = ΔC P / Δτ,
de ΔC S- change of molar concentration to the substrate (mol / l),
ΔC P- change of molar concentration to the reaction product (mol / l),
Δτ - hour (xv, sec).
Kinetic dosage is necessary to carry out at a saturated concentration of the substrate, in the case of a drop, the enzyme is not able to show maximum activity.
Single enzyme activity:
International Single Enzyme (U)- the amount of enzyme, which catalyzes the transformation of 1 micromol to the substrate for 1 quiline at a temperature of 25 ° C and the optimum pH of the middle.
In the CI system, one enzyme є katav (kat) This is a fraction of the enzyme, which catalyzes the transformation of one molar substrate in 1 second. It’s not important pidrahuvati, scho:
1 U = (1 * 10 -6 M) / 60 s = 1.67 * 10 -8 M s-1 = 1.67 * 10 -8 cat = 16.7 ncat.
often visit active activity the preparation in the enzyme increased the activity of the preparation of the enzyme, expressed in (U), on the basis of the preparation in milligrams:
A beats = U / mass to the drug (mg)
With purified enzymes in the pitoma, the activity increases. By the growth of pet activity, one can judge about the efficiency of the stages of purification and purity of the enzyme preparation.
For the assessment of the activity of highly purified, homogeneous preparations in enzymes, the number of international units (U) to the enzyme per enzyme per unit of speech (μmol) per enzyme rate molar activity(number of wraps). per physical zmist molar activity - the whole number of molecules to the substrate, which is transferred to one molecule of the enzyme for 1 quill or for 1 second. For example: for urease, accelerating the hydrolysis of sechovina, the molar activity becomes 30,000, trypsin - 102, glucose oxidase - 17,000 cycles per second.
the power of enzymes
4.1. Mechanism of the arts. Enzymes can change rapidly to catalyze reactions in the batch of products, in such a rank, the constant of effective reaction becomes permanent. Yak and all catalysts, ferments will not change the hour of reaching the price of rivnovagy. In a large number of enzymes, they accelerate reactions in 10 7 - 10 14 times. The efficiency of enzymatic catalysis is based on a strong decrease in the energy of activation of the reaction for the transformation of the substrate into the product through the transition plates.
4.2. specificity of the event... The specificity of the reaction with the substrate and the way through the enzymatic reaction begins with the apofenzyme. The specificity of the enzyme in the form of the conjugation of the exchange of speech in the organism.
It seems about enzymes, what a stink vuzka substrate specificity, There is a stench on the bow, there are few substrates. Inodi you can talk about absolutely substrate specificity, for example, catalase catalyzing only one reaction - dispensing with water peroxide:
Large enzymes are characterized by common (broad, group) Substrate specificity If the stench catalize a group of similar reactions. For example, alcohol dehydrogenase catalyzes the transformation of alcohols into aldehyde, and the substrates can be methanol, ethanol, propanol and other alcohols. This is the fact that alcohol dehydrogenase can oxidize non-linear alcohols, as well as an alcohol group, which can be included in the store of folding molecules, winter cream, and the enzyme can catalyze the transformation of retinol into retinal. Naturally, enzymes, over a wide substrate specificity, catalyze the transformation of substrates with a good efficiency.
Ferments of nadіlenі also stereochemical specificity: Їх active center develops molecules of substrates in a spacious configuration. For example, the oxidation of L-amino acids is active only as well as L-amino acids and absolutely does not exist on their D-analogues. For oxide deamination of D-amino acids in living organisms є oxidation of D-amino acids, but not on L-amino acids. The very fact that the active center is connected with the singular stereoisomers to the substrate lies in the basis of the function of such enzymes, such as racemazi, which transform one stereoisomer into one.
Specificity of the nobles of re-development However, one substrate from other enzymes can be transformed into products, which is developed for the structure and role in metabolism.
Guided butt: L-amino acid oxidase a diet on L-amino acids, converting them into alpha-keto acids with the solution of ammonia and peroxide water.
L-amino acid decarboxylase It is connected with these substrates, rather than catalyzing the reaction: decarboxylation from the formulations of biogenic amines and the form of carbon dioxide.
With just one butt є the possibility of converting glucose-6 phosphate from one of the other enzymes, one at a time from the various metabolic paths:
4.3. thermolability .
Yak and richly strong, when the temperature of the enzyme is adjusted, thermal denaturation is given, so that the native conformation of the enzyme is destroyed and the structure of the active center changes. Ferments of ssavts are repaired with denatured liquid at temperatures of 40 o C.
At a zyazyka z vyshekazanim, enzyme preparations are bazhano zberigati at low temperatures. One of the best ways to save enzymes is lyophilization (drying at temperatures lower than -70 ° C in vacuum), transferred to a partially denatured mill for additional salts of ammonia and placing in a refrigerator.
4.4. The prevalence of fluidity reactions in terms of temperature. The frequency of enzymatic reactions, such as any chemical reactions, is stored at a temperature. When the temperature is adjusted by 10 ° C, the speed of the reaction increases 2-4 times according to the Van't Hoff rule. However, at temperatures of 40 ° C, at the same stage of denaturation of enzymes, it is possible to produce to a decrease in the total activity (Fig. 2):
Small. 2. The presence of liquid enzymatic reactions in terms of temperature.
4.5. The severity of the fluidity of the reaction in terms of pH. The abundance of the liquid enzymatic reaction in the form of pH is bell-shaped (Fig. 3). The pH value, at which the most rapid enzymatic reaction occurs, is called optimal (pH-optimum). The nature of the curves and the value of the pH optimum is due to the nature of the charging of the groups to the substrate and the charging of the groups to the enzyme (especially quiet, as it enters the active center). The optimum pH for higher enzymes is between 6.0 and 8.0 (Fig. 3).
Small. 3. The severity of the liquid enzymatic reaction in terms of pH.
However, the єth vinyatka, for example, pepsin is most active at pH 1.5 - 2.0, and puddle phosphatase at pH 10.0 - 10.5 (Fig. 4)
Small. 4. Deposits of liquid enzymatic reactions (v) as the pH of the middle.
With extreme (even lower or even higher) pH values, the deterioration of the tertiary structure of the molecule is caused by the enzyme, which leads to the loss of enzymatic activity.
Similar information.
Understanding enzyme activity
In general biochemical practitioners, it is practical not to assess the amount of enzyme, but only its activity. Activity - more broad understanding, less number. The result of the reaction is the result of the reaction, and the recession itself to the substrate or the accumulation of the product. Naturally, at the same time, it is impossible to ignore an hour, if the enzyme is purified and the number of molecules to the enzyme. If the number of molecules for the enzyme to grow is unrealistic, then the amount of biological material is vicious to take revenge on the enzyme (obsyag abo masu).
With such a rank, when the activity of enzymes is significant, three changes are required one hour:
- the mass of the product of the darkened substrate;
- hour, staining for reaction;
- a bit of an enzyme, but for the sake of a lot of biological material, to take revenge on the enzyme.
For a clearer performance of significant factors with a simple and simple butt, there can be two budget servants. The budgets are primed to the product of the reaction, the work is done to the enzyme, the team will not help to discuss the biological material. Otzhe, zavdannya z 3rd class:
- There is a brigade of 10 people on the weekday of one worker, and a team of 5 people on the same day. The business ended overnight and generally. What is the activity of working people?
- A team of 10 people worked on the day of one booth from 3 surfaces, and a team of 10 people worked on the same day from 12 surfaces. The business ended overnight and generally. What is the activity of working people?
- A team of 10 people worked on the day of one booth from 5 surfaces, and a team of 10 people worked on the same day. The first week took 20 days, the other was inspired for 10 days. What is the activity of working people?
Fundamentals of enzyme activity values
1. The activity of the enzyme rotates in the liquidity of the accumulated product, or the liquidity of the substrate in the transfer to the amount of material, in order to take the enzyme.
Have a practitioner become vicious:
- single amount of speech - mol (і th thіdnі mmol, µmol), gram (kg, mg),
- at one hour - khvili, one hour, a second,
- odinitsі masi abo obsyagu - gram (kg, mg), liter (ml).
Actively perceptible and іnshі losses - katav (mol / s), the international unit of activity (ME, Unit) is given µmol / min.
Thus, the activity of the enzyme can vary, for example, in mmol / s × L, g / year × L, MO / L, cat / ml, etc.
For example, vidomo,
2. The standard mindset, the results can be adjusted, and the results can be adjusted in different laboratories - the optimal pH and the fixed temperature, for example, 25 ° C or 37 ° C, before the hour of incubation of the substrate with the enzyme.
enzymatic activity soil [from lat. Fermentum - ferment] -pdatn_st soil show a catalytic effect on the processes of transformation of exogenous and organic and mineral spoluks evident in n_y enzymes. Characterizing the enzymatic activity of soils, may be on the top of the total indicator of activity. The enzymatic activity of green soils is not identical and is linked to genetic features and a complex of interrelated ecology officials... The level of enzymatic activity of soils is due to the activity of enzymes (invertases, proteases, urease, dehydrogenases, catalase, phosphatases), which rotate through a number of unfolded substrate per 1 hour per 1 g
Biocatalytic activity of soils is found at the stage of enrichment in microorganisms and in the type of soil. The activity of enzymes varies according to genetic patterns, which are based on zyty humus, types of reactions, oxide potential and other indicators for the profile.
In long-leaved fossil soils, the intensity of enzymatic reactions is mainly in the horizon of the fossil layer, and in ornichi - orní shari. All biologically less active genetic horizons, which are located under the horizons A chi Ap, may have a low activity of enzymes. The activity of їх grows slightly when the soil is cultivated. The development of fossil soils before the growth of the enzymatic activity of the established forest horizon in the context of the fossilized soils lowers sharply, but in the world of new cultivation the soil tends to grow and become highly cultivated.
The enzymatic activity of the image of the state of the soil and internal changes, which is displayed in the case of the agricultural sector and the growing culture of agriculture. The change occurs when both long and fossil soils are introduced into the culture, as well as when the growth is growing.
For all Bilorus in ornichy soils, up to 0.9 t / ha of humus is usually consumed. As a result of the erosion, the humus rushes scrupulously and without rotation from the irrigation to 0.57 t / ha of humus. The reasons for the dehumification of soils є the improvement of the mineralization of the soil organic speech, The restoration of the processes of new humus as a result of mineralization in connection with the lack of goodness in the soil of organic goodness and a decrease in the enzymatic activity of the soil.
Biochemical transformation of organic speech in the soil is produced as a result of microbiological performance due to the injected enzymes. enzymatic activity of soil microorganism
A special role is played by enzymes in the living of tvarins, roslin and microorganisms. Soil ferments take on a role in the fall of grass, food and microbial surpluses, as well as the synthesis of humus. As a result of the vital speech, it is important to switch over to easily accessible forms for growing and microorganisms. Ferments are very active, very specific, and great fallowness. of different minds Zovnishny middle. The directors of the catalytic function of the stench will preserve the speed of projection in the body or the pose of the great number of chemical reactions.
Specially with the other criteria, the enzymatic activity of soils can serve as a superior diagnostic indicator for the improvement of the level of soil cultivation. As a result, doslidzhen 4, p. 91, the fallowness was established between the activity of microbial and enzymatic processes and the conducted visits, which increase the moisture of the soil. Processing of soils, introduced by goodness, changes the ecological situation and the development of microorganisms.
In a given hour, a few thousand of individual enzymes have been discovered in biological objects, and a few hundred of them have been seen and made. Vidomo scho Klitina is alive There can be up to 1000 different enzymes, which will accelerate this chemical reaction.
I am interested in the storage of enzymes in wickedness, and in the process of growing steadily in order to improve the safety of technological processes. Being present in our biological systems, being immediately products and instruments of our systems, enzymes are synthesized and function at physiological minds (pH, temperature, vice, presence of inorganic ions), when something is easy to get through. As a product, it also involves a lot of processes, it goes against the participation of enzymes, it is non-toxic and it is easy to handle. Moreover, in bagatokh vipadkah, ferments, which are vicarious in industry, are treated in an ecologically safe way. The type of non-biological catalysts of enzymes is not only safe and improved before biodegradation, but the specificity of the process, the way of thinking through the reactions of the high efficiency. Efficiency and specificity of enzymes allow you to cut off whole products in a high way, so that you can use ferments in industry in a cost-effective way. Stagnation of enzymes in the air, fast vitrati leads to energy in technological processes, changes in the atmosphere of CO2, reduces the risk of absorption dovkilla by-products of technological cycles.
The stagnation of advanced agricultural technology can change in the friendly direction of microbiological processes, not only of the one, but of the food ball in the soil.
For the non-mediated participation of late enzymes, the distribution of organic spoluk soil. Thus, proteolytic enzymes break down proteins into amino acids.
Urease has a storage capacity of up to CO2 and NH3. The statements of amiak and amoniyny salts serve as a dummy of nitrogen vitality of roslin and microorganisms.
Invertase and amylase take part in the cleavage in carbohydrates. The enzymes of the phosphate group lay down organophosphorus compounds in the soil and play an important role in the phosphate regime.
To characterize the enzymatic activity of the soil, use the most widespread enzyme, the power of the growth of the soil microflora - invertase, catalase, protease and іnshі.
In the minds of our republic there was held a minimum of doslidzhen 16, p. 115. Investigation of changes in the efficiency and enzymatic activity of soils during anthropogenic infusion, the protections of the data do not give a definite indication of the nature of changes through the folding of the results achieved in the sense of respect
In zv'yazku of CIM Poshuk optimal virіshennya problemi polіpshennya humus will ґruntu i її fermentativnoї aktivnostі in specific soil and klіmatichnih Minds on osnovі rozrobki resursozberіgayuchih priyomіv main obrobіtku pochvio zastosuvannya gruntozahisnih sіvozmіn scho spriyayut zberezhennyu structure zapobіgannya pereuschіlnennya ground i polіpshennya їh yakіsnogo that will renewal of soil fluidity in case of minimal vitrates, itself for an hour.
Doslidzhennya on the value of enzymatic activity MCD on the basis of small bacteria was carried out in the chotiri stage. To continue the boules, see the MKD studies based on the MKD-V monocultures (Bifidobacter bifidum longum), MKD-S ( Streptococcus termophilus), MKD-P (Propionobacterium acidi-propionicum), MKD-L (Lactobacillus acidophilus).
Meta doslidzhen is the value of building in probiotic microorganisms, which enter the warehouse of MKD, synthesize enzymes.
On the first stage according to GOST 20264.4-89 "Enzyme preparations. The method of determining the amylolytic activity "in all patients of the MCD by the Ison method, the total amylolytic activity began. Anson's method of storing on starch hydrolysis with an enzyme-amylolytic complex to dextrin in a molecular weight.
On another stage according to GOST 20264.2-88 “Enzyme preparations. The method of evaluating proteolytic activity "started the total proteolytic activity (PA) in pre-juvenile stages. The method of inoculation on hydrolysis of sodium caseinate with an enzyme complex at pH-7.2. For the value of PA neutral protease 0-10 u / ml, change the minimum dilution. Acid protease started at pH-5.5, when hydrolysis of the protein passed through.
At the third stage According to TU9291-008-13684916-05, in all presentations of the MCD, the beginning of the CELLULOSOLITIC activity (CA) began. The method of cellulase detection is based on the designation of the synthesis of sugar as a result of cellulase hydrolysis and chromatographic paper on the enzyme. The method of recommendations by the International Committee of the IUPAC on biotechnology as the main test for cellulase activity. For the unit of cellulase efficiency, take the same amount of enzyme, as for chromatographic papier at 50 ° C and pH 4.8, the concentration is 1 mmol. As a rule, the activity rotates in units per mil.
ostanim, fourth, stage increased activity of lipase (LA). The method of assigning lipase to the Skerman technique of saturating fatty acids on a titrated meadow, which was adopted for a second lipase, when vicorian in the quality of olive oil substrate. In case of a significant lipase, the reaction is complete, it should be stored in 6.5 ml of 1/15 M phosphate-citrate buffer, 2.5 ml of an olive oil emulsion in a 1% solution of polyvinyl alcohol in alcohol: 3 ml ridini.
Previous MCDs based on different microorganisms-probiotics showed that all prior MCDs could replace one or more groups of enzymes (Table 8). Thus, MKD-V showed the presence of all pre-juvenile groups of enzymes. MKD-R to place three groups of enzymes in its warehouse: amylolytic, proteolytic, cellulolytic. In MKD-L, there are also three groups of enzymes: proteolytic, cellulolytic and lipolytic, the amylolytic group is weakly expressed. MKD-S has two groups of enzymes in its warehouse: proteolytic and cellulose, amylolytic and lypolytic activity of the turnover is weak.
Table 8
Enzymatic activity of MKD, od / ml
The analysis of the data, rejected as a result of the pretreatment of the enzymatic activity of MKD, may be in high step cellulase activity - from 64.46 (MKD-R) to 72.4 IU / ml (MKD-L).
MKD-S and MKD-V may have the same activity of cellulase - 66.7 u / ml.
All MCDs have been allowed to replace acidic protease, in which hydrolysis the protein is healthy at pH-5.5. The highest activity of protease is revealed in MKD-R - 7.5 u / ml, namensha - in MKD-L (1.0 u / ml). MKD-B value of protease activity 2.0, MKD-S - 2.5 u / ml.
Lipase activity is indicated in MKD-L - 1.4 and in MKD-V - 12.6 u / ml. In MKD-S and MKD-R, the lipase enzyme is not recognized.
Amilolytic activity is revealed in MKD-V - 11.2 and in MKD-R - 9.4 u / ml.
In fig. 1 ^ 1 graphically presents the value of the enzymatic activity of MKD.
Small. 1.
Small. 2.
Small. 3.
Small. 4.
The results of the preliminarily enzymatic activity of MCD on the basis of young bacteria are used in literature data about those, that bacteria-probation-volodiut enzymatic activity. Our preliminaries discovered the enzyme powers of bifidobacteria at the MKD warehouse in some cases with pre-sensuous microorganisms. According to the data, up to 90% of the representatives of the normal flora of the intestines of birds are stored in bifidobacteria. Bifidobacteria are found in all intestinal tracts.
Enzyme groups synthesized by them take part in all enzymatic processes during the re-transformation of lively vernacular fodder in the intestinal tract. On A. І. Havkina (2003), bifidobacteria take an active part in the processes of enzyme pre-digestion of feeds, while feeding on hydrolysis proteins, fermenting in carbohydrates, absorbing fat, and eliminating the enzyme steps in the cells of all storage Otrimanі danі inform about the specificity іvnі and spectrum of fermented enzyme groups in fallowness from belonging to microorganisms to singing species and minds of living. Due to the fact that due to their higher characteristics, we are guilty of the development of new recommendations for the supply of quiet types of probiotic feed additives.
The enzymatic activity of microorganisms in the bagat is also versatile. According to them, it is possible to establish not only the species and the type of belonging of the microbe, but also the significance of its options (so called biovari). The basic enzymatic power and quality are easily discernible.
cleavage in carbohydrates(Sugarolytic activity), i.e., the healthiness of degradation of sugar and rich atomic alcohol from acid and acid and gas solutions, injected in the middle of the Giss, as it is to replace that one carbohydrate and indicator. When the acid is cleaved in the carbohydrate, it is an indicator of the change in the middle. The middle course is called "row row". Mikrobi, which does not ferment Danium into carbohydrates, grow in the middle, do not change. Evidently, gas will be installed for lighting the bulb in the middle with agar, or after bought it in the "floats" in the middle of the middle. "Float" - a vuzka vuzka a tube with a sealed end, burnt into the mountain, before sterilization is placed in a test tube with the middle (Fig. 18).
Small. 18. Vivchennya saccharolytic activity of microorganisms. I - "row row": a - middle row with carbohydrates і indicator Andrede; b - for example, the middle with the BP indicator: 1 - microorganisms do not ferment into the carbohydrate; 2 - microorganisms ferment into carbohydrate with acid; 3 - microorganisms ferment into carbohydrates with acid and gas solutions; II - colonies of microorganisms, which do not deposit (without barriers) and deposit lactose (violets in the middle of the EMC - zliv, chervons in the middle Endo - right-handed)
In addition, the saccharolytic activity is vivchayut in the middle of Endo, EMC, Ploskirєva. Microorganisms, fermenting to acidity, are found in the middle of the milk tsukor (lactose), fix the colony - the acid of the change in the color of the obvious in the middle of the indicator. Colonies of microbes, which do not ferment lactose, without barn (div. Fig. 18).
Milk with growth of microbes, fermenting lactose, burns out.
With the growth of microorganisms, where amylase is set up, in the middle of the starch with a small starch, the splitting occurs. You know about the price, having brought Lugol's line to the culture of the splinter - the color of the middle does not change. Non-cleavage of starch is even due to the size of the blue barn.
proteolytic power(T.E. Health of spitting up beads, polypeptides, etc.) Vivchayut in the middle with gelatin, milk, syrovatka, peptone. When growing on the gelatinous middle of the microbes, ferment the gelatin, the middle grows. The nature of the growth, how to get caught up in small microbes, grows (Fig. 19). Mikrobi, which dissolves casein (milk beetle), viklikayut milk peptonezatsiya - vono swelling milk syrovat. When peptones are split, indol, syrwater, amiak can be seen. Їх will be installed for the additional help of indicator papіrtsіv. The filter papier is smelled out of the way with singing ropes, hanging, dividing with vuzhny pigs up to 5-6 cm and when sowing the culture on the BCH to help the cork between her and the test tube. Pislya incubation in thermostats will provide the result. Amiak wiklikak blue litmus papirtsya; if water is seen on papyrtsi, leaked with 20% strength to lead acetate and sodium hydrogen carbonate, the approval of lead sulphate - papyretsya chorniyu is taken; Indol viklikє pochvonіnnya papіrtsі, leaked with a large amount of oxalic acid (div. Fig. 19).
Small. 19. Proteolytic power of microorganisms. 1 - form razrіdzhennya gelatin; II - viznachennya sirkovodnyu; III - value of indole: 1 - negative result; 2 - positive result
In addition to the meanings of the middle, the building of microorganisms in the development of growing substrates is used for the supplementary paper disks, leaked with the help of reagents (system of indicators of paper "SIB"). These disks are lowered into tubes with pre-juvenile culture, and after 3 years incubation in thermostats at 37 ° C, according to the change in the color of the disks, one can judge about the dispensed in carbohydrates, amino acids, bile acids, etc.
Hemolytic power (health of ruinuvati erythrocytes) vivchayut in the middle with blood. Ridky middle ground at the same time will be clear-cut, and on the middle middle ground near the colony there will be a hole zone (Fig. 20). When methemoglobin is approved, the middle is green.
Small. 20. Hemolysis near colonies to grow on blood agar
preserving cultures
Vidіlenі і vivchenі cultures (shtami), which represent value for science, or vyrobnіstva, take in museums of living cultures. The All-Union Museum is located in the Sovereign NDI Standardization and Control of Medical Biological Preparations. L.A. Tarasevich (GISK).
The head of the zberigannya is to educate the life of microorganisms and to get ahead of their ministry. For a wide demand, weaken or pinch exchange in microbial cells.
One of the most thorough methods of trivial preservation of cultures is lyophilization - hanging in a vacuum from a frozen mill, allowing the setting of anabiosis. Visushuvannya to spend in special apparatuses. Take culture in sealed ampoules at a temperature of 4 ° C, more beautifully at -30-70 ° C.
Renovation of hanging crops. Strongly heat the tip of the ampoule in the half of the pallet and push it up with a cotton swab, slightly soaked with cold water, microscopic spots appear on the slopes, through which it is easy to seep into the middle of the ampoule. At the same time, passing through the rosіgrіtі land of thrusts, wіth wіth thе sterilization.
* (If there is too much water on the tampon, you can drink it into the ampoule and destroy the sterility of the culture: її is soaked through the microscopes, so there’s a vacuum in the ampoule.)
Uwaga! Do not forget that there is a vacuum in the sealed ampoule. As soon as you drink in it, you drink it all at once through the great opening, you can sprinkle it in the ampoule of culture and status of the Wikid.
If you let go by chance, I quickly smack and see the top of the ampoule with a pair of tweezers. Slightly scorch the openings with a sterile pasteurized pipette, or add a syringe to the ampoule with a bottle (broth or isotonic). Remix ampoules instead of and transfer to the middle. The growth of new crops in the first crops can be made.
Trivially zberigati culture is also possible in natural nitrogen (-196 ° C) in special attachments.
Methods of non-trivial preservation of cultures of the offensive: 1) subculture (periodical repetition in the middle of the middle) with intervals, so that the powers of microorganism, middle and minds of cultivation can be found. If I re-feed the culture, take it at 4 ° C; 2) saving a ball of butter. Grow the culture in 5-6 cm agar, pour sterile vaseline oil (a ball of oil about 2 cm) and store it vertically in the refrigerator. The term is taken from the small microorganisms of the development, to that from the test tubes periodically hang culture, to reconsider the life; 3) zberigannya at -20-70 ° C; 4) zberigannya in sealed test tubes. If necessary, take the material hanging on the middle.
Control food
1. What to enter into the understanding "bacteriological doslіdzhennya"?
2. What kind of culture is there for such an experience?
3. What is the colony of microbes, culture, strain, clone?
4. How to enter into the understanding of "cultural power of bacteria"?
zavdannya
1. Vivchit and describe the number of colonies. Transfer to the agar mows and to the sector.
2. Vivchit and describe the nature of growth - culture on slant agar. Look for the purity and morphology of the culture in the prepared preparation.
3. Perez culture from slant agar on broth and on differential diagnostics middle. Vivchit and write down in the protocol the nature of the growth of culture in the midst and the enzymatic power.