提高双歧杆菌活菌制剂常温贮存稳定性的实验研究

以Ｄ８５０４青春双歧杆菌为实验３菌株,对活菌制剂常温保存的稳定性问题探讨。首先对菌种进行耐温耐氧驯化试验,然后进行双歧杆菌菌体冻干过程中加入适宜保护剂试验。结果双歧杆菌活菌制剂常温保存的稳定性显著提高。经常温贮存２７０ｄ后检测,质量符合规定。     

双歧酸奶定量干燥菌种的实验室研究

目的:对双歧酸奶定量干燥菌种的实验室研究进行初步探讨.方法:培养收获三种乳酸菌(双歧杆菌、乳杆菌、嗜热链球菌),并采用真空冷冻干燥技术进行处理,从而对干燥效果以及常温保存活性进行评价和分析.结果:三种乳酸菌的干燥菌粉活菌计数均大于1012个/g;凝乳试验显示次代菌种发酵时间明显低于第一代;混合发酵的凝乳中活菌计数均大于106个/ml,单一菌种发酵的凝乳中活菌计数超过1010个/ml;干燥菌种在37℃放置30 d后,活菌数仍可达108个/g以上.结论:双歧酸奶定量干燥菌种实验室研究结果令人满意,为开发双歧酸奶袋装式干燥菌种打下了基础.     

双歧杆菌活菌制剂治疗烧伤腹泻的临床观察与分析

本研究采用双歧杆菌活菌制剂治疗烧伤后腹泻病人２６例,发现腹泻时患者肠道菌群发生明显变化：总菌量下降,厌氧菌中的双歧杆菌、类杆菌的数量明显减少,念珠菌的数量明显升高,厌氧菌与需氧菌比例下降,发生菌群失调。喂服双歧杆菌活菌制剂后,双歧杆菌数量增加,需氧、厌氧菌比例下降,患者失调的肠道菌群趋向正常,腹泻停止。双歧杆菌的服用重建了肠道菌群微生态平衡,达到治疗腹泻的目的     

青春双歧杆菌口嚼片生产中双歧杆菌存活因素初探

对生产双歧杆菌口嚼片过程中影响双歧杆菌存活的部分因素进行了研究,考察菌龄、干燥制剂的配料中辅应剂成分、水分含量等影响双歧杆菌稳定性的因素。结果表明,配料中一定比例的棕榈油、乙基纤维等疏水性填充剂具有保护与稳定双歧杆菌存活的作用,在双歧杆菌发酵液中,采用对数生长末期的细胞制备的干燥粉料中活菌数及其存活率都较高。产品水分含量以５％～６％、充氮包装、存放于低温环境更有利于提高存活率。     

双歧豆奶菌种的驯化研究

选用青春双歧杆菌在 A、B、C三种培养基上进行耐氧和凝固驯化 ,经 2 8代筛选出一株耐氧性并在 C培养基上进行耐氧和凝固驯化 ,经 2 8代筛选出一株耐氧性并在 C培养基中由 5 6 h可发生凝固 ,最后缩短到 12 h发生凝固的青春双歧杆菌 ;在 PYG和豆乳培养基中进行耐酸驯化 ,经 15代筛出一株能在p H4.7°T90的发酵豆乳中存活的青春双歧杆菌 ;在 BCPYC液体和固体培养基中进行耐热驯化 ,经 14代筛选出一株能在 47℃± 1℃的环境中生长的青春双歧杆菌。本菌在改良的 NPNL双歧选择培养基中生长 ,形态、培养与生化特性与原菌种无差异。     

人类肠道双歧杆菌一新种的初步研究

在健康人群肠道双歧杆菌检索中,从中老年粪便先后分离出一种细胞形态呈环状双歧杆菌３株（编号为ＭＯ－５,ＭＯ－２及ＭＯ－１）。经生物学特征的常规鉴定,根据《伯杰系统细菌手册》第２卷１９８６＾［２］。及其他有关文献描述,该菌的生理生化特征与双歧杆菌属中的青春双歧杆菌相类似,但从该菌的细胞形态呈环状和水杨苷发酵反应的阴性结果,再与青春双歧杆菌相比有区别。故认为该菌可能归属于双歧杆菌属中一新种。暂称它为环状     

酪酸梭菌-婴儿型双歧杆菌二联活菌制剂的研究

对酪酸梭菌－婴儿型双歧杆菌二联活菌制剂的特性及功效进行研究和分析。方法：通过实验鼠进行刺激生长试验、活菌数测定、毒理试验、调节肠道菌群试验、免疫调节作用来观察制剂的作用。结果：小鼠肠道中双歧杆菌、乳杆菌、酪酸梭菌数量明显增加,高剂量能增加小鼠抗体生成细胞数。     

双歧杆菌四联活菌片中嗜酸乳杆菌的分离及生长动力学研究

目的以双歧杆菌四联活菌片为实验材料,利用酸化的MRS培养基筛选分离得到嗜酸乳杆菌,对其进行进一步的生长动力学研究,确定嗜酸乳杆菌的生长数学模型。方法通过浓度梯度稀释法利用改良MRS培养基对双歧杆菌四联活菌片中的嗜酸乳杆菌进行分离,利用分光光度仪和平板菌落计数两种不同的方法测定嗜酸乳杆菌在发酵过程中不同发酵时间的细胞浓度的动态变化,经软件处理后拟合出嗜酸乳杆菌细胞生长的Logistic数学模型。结果Logistic方程能很好地拟合嗜酸乳杆菌细胞生长的动态变化,并得到嗜酸乳杆菌在本实验条件下的数学模型,为进一步研究、利用嗜酸乳杆菌生长能力、产酸能力和产香能力等具有重要的理论指导意义。     

灭活的双歧杆菌对EPEC的黏附抑制作用

目的：研究灭活的青春双歧杆菌DMS8504对肠致病灶大肠埃希菌（EPEC）黏附抑制作用。方法：通过与活菌比较,观察灭活的双歧杆菌粘附于人大肠癌CCL－229细胞后对EPEC的黏附抑制作用。结果：用SCS或pH5.0新鲜BS肉汤悬浮的双歧杆菌能够安全抑制EPEC的黏附,而仅用SCS或pH5.0新鲜BS肉汤均不能抑制其黏附。     

青春双歧杆菌的波状层次生长特征观察

通过观察青春双歧杆菌在体外培养条件出现波状层次生长的情况,了解该过程中青春双歧杆菌形态的改变及其生物学意义。采用青春双歧杆菌低浓度接种于BS固体平板培养基中,延长培养时间至6 d,使其单个菌落长成较大的菌落后,置4℃冰箱中2~4周,在其培养结束时和放置冰箱后对菌落及菌体形态特征进行观察。在特定的培养时间及其后的低温处理后,青春双歧杆菌部分菌落出现了半透明环和不透明环交替的层次状结构,在半透明环区域和不透明环区域,菌体的形态以及对温度的敏感性存在差异,且在透明环区域,菌体发生了自溶现象。证明青春双歧杆菌在一定条件下形成了波状层次生长过程,且在该过程中,青春双歧杆菌的生长、生存方式以及增殖方式均发生了适应性变化。     

THE EFFECT OF ACETATE BUFFER MIXTURES,ACETIC ACID,AND SODIUM ACETATE,ON THE PROTOPLASM,AS INFLUENCING THE RATE OF PENETRATION OF CRESYL BLUE INTO THE VACUOLE OF NITELLA

When living cells of Nitella are exposed to a solution of sodium acetate and are then placed in a solution of brilliant cresyl blue made up with a borate buffer mixture at pH 7.85, a decrease in the rate of penetration of dye is found, without any change in the pH value of the sap. It is assumed that this inhibiting effect is caused by the action of sodium on the protoplasm. This effect is not manifest if the dye solution is made up with phosphate buffer mixture at pH 7.85. It is assumed that this is due to the presence of a greater concentration of base cations in the phosphate buffer mixture. In the case of cells previously exposed to solutions of acetic acid the rate of penetration of dye decreases with the lowering of the pH value of the sap. This inhibiting effect is assumed to be due chiefly to the action of acetic acid on the protoplasm, provided the pH value of the external acetic acid is not so low as to involve an inhibiting effect on the protoplasm by hydrogen ions as well. It is assumed that the acetic acid either has a specific effect on the protoplasm or enters as undissociated molecules and by subsequent dissociation lowers the pH value of the protoplasm. With acetate buffer mixture the inhibiting effect is due to the action of sodium and acetic acid on the protoplasm. The inhibiting effect of acetic acid and acetate buffer mixture is manifested whether the dye solution is made up with borate or phosphate buffer mixture at pH 7.85. It is assumed that acetic acid in the vacuole serves as a reservoir so that during the experiment the inhibiting effect still persists.     

AMPHOTERIC COLLOIDS : V. THE INFLUENCE OF THE VALENCY OF ANIONS UPON THE PHYSICAL PROPERTIES OF GELATIN.

1. When we plot the values of osmotic pressure, swelling, and viscosity of gelatin solutions as ordinates over the pH as abscissæ, practically identical curves are obtained for the effect of monobasic acids (HCl, HBr, HNO₃, and acetic acid) on these properties. 2. The curves obtained for the effect of H₂SO₄ on gelatin are much lower than those obtained for the effect of monobasic acids, the ratio of maximal osmotic pressures of a 1 per cent solution of gelatin sulfate and gelatin bromide being about 3:8. The same ratio had been found for the ratio of maximal osmotic pressures of calcium and sodium gelatinate. 3. The curves representing the influence of other dibasic and tribasic acids, viz. oxalic, tartaric, succinic, citric, and phosphoric, upon gelatin are almost identical with those representing the effect of monobasic acids. 4. The facts mentioned under (2) and (3) permit us to decide between a purely chemical and a colloidal explanation of the influence of acids on the physical properties of gelatin. In the former case we should be able to prove, first, that twice as many molecules of HBr as of H₂SO₄ combine with a given mass of gelatin; and, second, that the same number of molecules of phosphoric, citric, oxalic, tartaric, and succinic acids as of HNO₃ or HCl combine with the same mass of gelatin. It is shown in the present paper that this is actually the case. 5. It is shown that gelatin sulfate and gelatin bromide solutions of the same pH have practically the same conductivity. This disproves the assumption of colloid chemists that the difference in the effect of bromides and sulfates on the physical properties of gelatin is due to a different ionizing and hydratating effect of the two acids upon the protein molecule.     

ON THE INFLUENCE OF AGGREGATES ON THE MEMBRANE POTENTIALS AND THE OSMOTIC PRESSURE OF PROTEIN SOLUTIONS

1. It is shown that when part of the gelatin in a solution of gelatin chloride is replaced by particles of powdered gelatin (without change of pH) the membrane potential of the solution is influenced comparatively little. 2. A measurement of the hydrogen ion concentration of the gelatin chloride solution and the outside aqueous solution with which the gelatin solution is in osmotic equilibrium, shows that the membrane potential can be calculated from this difference of hydrogen ion concentration with an accuracy of half a millivolt. This proves that the membrane potential is due to the establishment of a membrane equilibrium and that the powdered particles participate in this membrane equilibrium. 3. It is shown that a Donnan equilibrium is established between powdered particles of gelatin chloride and not too strong a solution of gelatin chloride. This is due to the fact that the powdered gelatin particles may be considered as a solid solution of gelatin with a higher concentration than that of the weak gelatin solution in which they are suspended. It follows from the theory of membrane equilibria that this difference in concentration of protein ions must give rise to potential differences between the solid particles and the weaker gelatin solution. 4. The writer had shown previously that when the gelatin in a solution of gelatin chloride is replaced by powdered gelatin (without a change in pH), the osmotic pressure of the solution is lowered the more the more dissolved gelatin is replaced by powdered gelatin. It is therefore obvious that the powdered particles of gelatin do not participate in the osmotic pressure of the solution in spite of the fact that they participate in the establishment of the Donnan equilibrium and in the membrane potentials. 5. This paradoxical phenomenon finds its explanation in the fact that as a consequence of the participation of each particle in the Donnan equilibrium, a special osmotic pressure is set up in each individual particle of powdered gelatin which leads to a swelling of that particle, and this osmotic pressure is measured by the increase in the cohesion pressure of the powdered particles required to balance the osmotic pressure inside each particle. 6. In a mixture of protein in solution and powdered protein (or protein micellæ) we have therefore two kinds of osmotic pressure, the hydrostatic pressure of the protein which is in true solution, and the cohesion pressure of the aggregates. Since only the former is noticeable in the hydrostatic pressure which serves as a measure of the osmotic pressure of a solution, it is clear why the osmotic pressure of a protein solution must be diminished when part of the protein in true solution is replaced by aggregates.     

THE COLLOIDAL BEHAVIOR OF PROTEINS

1. It is well known that neutral salts depress the osmotic pressure, swelling, and viscosity of protein-acid salts. Measurements of the P.D. between gelatin chloride solutions contained in a collodion bag and an outside aqueous solution show that the salt depresses the P.D. in the same proportion as it depresses the osmotic pressure of the gelatin chloride solution. 2. Measurements of the hydrogen ion concentration inside the gelatin chloride solution and in the outside aqueous solution show that the difference in pH of the two solutions allows us to calculate the P.D. quantitatively on the basis of the Nernst formula See PDF for Equation if we assume that the P.D. is due to a difference in the hydrogen ion concentration on the two sides of the membrane. 3. This difference in pH inside minus pH outside solution seems to be the consequence of the Donnan membrane equilibrium, which only supposes that one of the ions in solution cannot diffuse through the membrane. It is immaterial for this equilibrium whether the non-diffusible ion is a crystalloid or a colloid. 4. When acid is added to isoelectric gelatin the osmotic pressure rises at first with increasing hydrogen ion concentration, reaches a maximum at pH 3.5, and then falls again with further fall of the pH. It is shown that the P.D. of the gelatin chloride solution shows the same variation with the pH (except that it reaches its maximum at pH of about 3.9) and that the P.D. can be calculated from the difference of pH inside minus pH outside on the basis of Nernst''s formula. 5. It was found in preceding papers that the osmotic pressure of gelatin sulfate solutions is only about one-half of that of gelatin chloride or gelatin phosphate solutions of the same pH and the same concentration of originally isoelectric gelatin; and that the osmotic pressure of gelatin oxalate solutions is almost but not quite the same as that of the gelatin chloride solutions of the same pH and concentration of originally isoelectric gelatin. It was found that the curves for the values for P.D. of these four gelatin salts are parallel to the curves of their osmotic pressure and that the values for pH inside minus pH outside multiplied by 58 give approximately the millivolts of these P.D. In this preliminary note only the influence of the concentration of the hydrogen ions on the P.D. has been taken into consideration. In the fuller paper, which is to follow, the possible influence of the concentration of the anions on this quantity will have to be discussed.     

AMPHOTERIC COLLOIDS : III. CHEMICAL BASIS OF THE INFLUENCE OF ACID UPON THE PHYSICAL PROPERTIES OF GELATIN.

1. The method of removing the excess of hydrobromic acid after it has had a chance to react chemically with gelatin has permitted us to measure the amount of Br in combination with the gelatin. It is shown that the curves representing the amount of bromine bound by the gelatin are approximately parallel with the curves for the osmotic pressure, the viscosity, and swelling of the gelatin solution. This proves that the curves for osmotic pressure are an unequivocal function of the number of gelatin bromide molecules formed under the influence of the acid. The cc. of 0.01 N Br in combination with 0.25 gm, of gelatin we call the bromine number. 2. The explanation of this influence of the acid on the physical properties of gelatin is based on the fact that gelatin is an amphoteric electrolyte, which at its isoelectric point is but sparingly soluble in water, while its transformation into a salt with a univalent anion like gelatin Br makes it soluble. The curve for the bromine number thus becomes at the same time the numerical expression for the number of gelatin molecules rendered soluble, and hence the curve for osmotic pressure must of necessity be parallel to the curve for the bromine number. 3. Volumetric analysis shows that gelatin treated previously with HBr is free from Br at the isoelectric point as well as on the more alkaline side from the isoelectric point (pH ≧ 4.7) of gelatin. This is in harmony with the fact that gelatin (like any other amphoteric electrolyte) can dissociate on the alkaline side of its isoelectric point only as an anion. On the more acid side from the isoelectric point gelatin is found to be in combination with Br and the Br number rises with the pH. 4. When we titrate gelatin, treated previously with HBr but possessing a pH = 4,7, with NaOH we find that 25 cc. of a 1 per cent solution of isoelectric gelatin require about 5.25 to 5.5 cc. of 0.01 N NaOH for neutralization (with phenolphthalein as an indicator). This value which was found invariably is therefore a constant which we designate as "NaOH (isoelectric)." When we titrate 0.25 gm. of gelatin previously treated with HBr but possessing a pH < 4.7 more than 5.5 cc. of 0.01 N NaOH are required for neutralization. We will designate this value of NaOH as "(NaOH)_n," where n represents the value of pH. If we designate the bromine number for the same pH as "Br_n" then we can show that the following equation is generally true: (NaOH)_n = NaOH (isoelectric) + Br_n. In other words, titration with NaOH of gelatin (previously treated with HBr) and being on the acid side of its isoelectric point results in the neutralization of the pure gelatin (NaOH isoelectric) with NaOH and besides in the neutralization of the HBr in combination with the gelatin. This HBr is set free as soon as through the addition of the NaOH the pH of the gelatin solution becomes equal to 4.7. 5. A comparison between the pH values and the bromine numbers found shows that over 90 per cent of the bromine or HBr found was in our experiments in combination with the gelatin.     

HYDRATION OF GELATIN IN SOLUTION

1. It was shown that the high viscosity of gelatin solutions as well as the character of the osmotic pressure-concentration curves indicates that gelatin is hydrated even at temperatures as high as 50°C. 2. The degree of hydration of gelatin was determined by means of viscosity measurements through the application of the formula See PDF for Equation. 3. When the concentration of gelatin was corrected for the volume of water of hydration as obtained from the viscosity measurements, the relation between the osmotic pressure of various concentrations of gelatin and the corrected concentrations became linear, thus making it possible to determine the apparent molecular weight of gelatin through the application of van''t Hoff''s law. The molecular weight of gelatin at 35°C. proved to be 61,500. 4. A study was made of the mechanism of hydration of gelatin and it was shown that the experimental data agree with the theory that the hydration of gelatin is a pure osmotic pressure phenomenon brought about by the presence in gelatin of a number of insoluble micellæ containing a definite amount of a soluble ingredient of gelatin. As long as there is a difference in the osmotic pressure between the inside of the micellæ and the outside gelatin solution the micellæ swell until an equilibrium is established at which the osmotic pressure inside of the micellæ is balanced by the total osmotic pressure of the gelatin solution and by the elasticity pressure of the micellæ. 5. On addition of HCl to isoelectric gelatin the total activity of ions inside of the micellæ is greater than in the outside solution due to a greater concentration of protein in the micellæ. This brings about a further swelling of the micellæ until a Donnan equilibrium is established in the ion distribution accompanied by an equilibrium in the osmotic pressure. Through the application of the theory developed here it was possible actually to calculate the osmotic pressure difference between the inside of the micellæ and the outside solution which was brought about by the difference in the ion distribution. 6. According to the same theory the effect of pH on viscosity of gelatin should diminish with increase in concentration of gelatin, since the difference in the concentration of the protein inside and outside of the micellæ also decreases. This was confirmed experimentally. At concentrations above 8 gm. per 100 gm. of H₂O there is very little difference in the viscosity of gelatin of various pH as compared with that of isoelectric gelatin.     

DONNAN EQUILIBRIUM AND THE PHYSICAL PROPERTIES OF PROTEINS : II. OSMOTIC PRESSURE.

1. It had been shown in previous publications that the osmotic pressure of a 1 per cent solution of a protein-acid salt varies in a characteristic way with the hydrogen ion concentration of the solution, the osmotic pressure having a minimum at the isoelectric point, rising steeply with a decrease in pH until a maximum is reached at pH of 3.4 or 3.5 (in the case of gelatin and crystalline egg albumin), this maximum being followed by a steep drop in the osmotic pressure with a further decrease in the pH of the gelatin or albumin solution. In this paper it is shown that (aside from two minor discrepancies) we can calculate this effect of the pH on the osmotic pressure of a protein-acid salt by assuming that the pH effect is due to that unequal distribution of crystalloidal ions (in particular free acid) on both sides of the membrane which Donnan''s theory of membrane equilibrium demands. 2. It had been shown in preceding papers that only the valency but not the nature of the ion (aside from its valency) with which a protein is in combination has any effect upon the osmotic pressure of the solution of the protein; and that the osmotic pressure of a gelatin-acid salt with a monovalent anion (e.g. Cl, NO₃, acetate, H₂PO₄, HC₂O₄, etc.) is about twice or perhaps a trifle more than twice as high as the osmotic pressure of gelatin sulfate where the anion is bivalent; assuming that the pH and gelatin concentrations of all the solutions are the same. It is shown in this paper that we can calculate with a fair degree of accuracy this valency effect on the assumption that it is due to the influence of the valency of the anion of a gelatin-acid salt on that relative distribution of the free acid on both sides of the membrane which Donnan''s theory of membrane equilibrium demands. 3. The curves of the observed values of the osmotic pressure show two constant minor deviations from the curves of the calculated osmotic pressure. One of these deviations consists in the fact that the values of the ascending branch of the calculated curves are lower than the corresponding values in the curves for the observed osmotic pressure, and the other deviation consists in the fact that the drop in the curves of calculated values occurs at a lower pH than the drop in the curves of the observed values.     

THE ISOELECTRIC POINT OF A STANDARD GELATIN PREPARATION1

Two samples of a standard gelatin were studied, both prepared according to published specifications and washed free from diffusible electrolytes. The isoelectric point of this material was determined in four ways. 1. The pH values of solutions of gelatin in water approached the limit 4.86 ± 0.01 as the concentration of gelatin was increased. 2. The pH values of acetate buffers were unchanged by the addition of gelatin only at pH 4.85 ± 0.01. This gives the isoionic point of Sørensen, which is the isoelectric point with respect only to hydrogen and hydroxyl ions. 3. Gels of this gelatin made up in dilute HCl or NaOH, or in dilute acetate buffers, exhibited maximum turbidity at pH 4.85 ± 0.03. 4. Very dilute suspensions of collodion particles in 0.1 per cent gelatin solutions made up in acetate buffers showed zero velocity in cataphoresis experiments only at pH 4.80 ± 0.01. No evidence was found for the assumption that gelatin has two isoelectric points at widely separated pH values. It is concluded that the isoelectric point of this standard gelatin is not far from pH 4.85.     

AMPHOTERIC COLLOIDS : II. VOLUMETRIC ANALYSIS OF ION-PROTEIN COMPOUNDS; THE SIGNIFICANCE OF THE ISOELECTRIC POINT FOR THE PURIFICATION OF AMPHOTERIC COLLOIDS.

1. It is shown by volumetric analysis that on the alkaline side from its isoelectric point gelatin combines with cations only, but not with anions; that on the more acid side from its isoelectric point it combines only with anions but not with cations; and that at the isoelectric point, pH = 4.7, it combines with neither anion nor cation. This confirms our statement made in a previous paper that gelatin can exist only as an anion on the alkaline side from its isoelectric point and only as a cation on the more acid side of its isoelectric point, and practically as neither anion nor cation at the isoelectric point. 2. Since at the isoelectric point gelatin (and probably amphoteric colloids generally) must give off any ion with which it was combined, the simplest method of obtaining amphoteric colloids approximately free from ionogenic impurities would seem to consist in bringing them to the hydrogen ion concentration characteristic of their isoelectric point (i.e., at which they migrate neither to the cathode nor anode of an electric field). 3. It is shown by volumetric analysis that when gelatin is in combination with a monovalent ion (Ag, Br, CNS), the curve representing the amount of ion-gelatin formed is approximately parallel to the curve for swelling, osmotic pressure, and viscosity. This fact proves that the influence of ions upon these properties is determined by the chemical or stoichiometrical and not by the "colloidal" condition of gelatin. 4. The sharp drop of these curves at the isoelectric point finds its explanation in an equal drop of the water solubility of pure gelatin, which is proved by the formation of a precipitate. It is not yet possible to state whether this drop of the solubility is merely due to lack of ionization of the gelatin or also to the formation of an insoluble tautomeric or polymeric compound of gelatin at the isoelectric point. 5. On account of this sudden drop slight changes in the hydrogen ion concentration have a considerably greater chemical and physical effect in the region of the isoelectric point than at some distance from this point. This fact may be of biological significance since a number of amphoteric colloids in the body seem to have their isoelectric point inside the range of the normal variation of the hydrogen ion concentration of blood, lymph, or cell sap. 6. Our experiments show that while a slight change in the hydrogen ion concentration increases the water solubility of gelatin near the isoelectric point, no increase in the solubility can be produced by treating gelatin at the isoelectric point with any other kind of monovalent or polyvalent ion; a fact apparently not in harmony with the adsorption theory of colloids, but in harmony with a chemical conception of proteins.     

FRACTIONATION OF GELATIN

1. It is possible to fractionate gelatin by means of reprecipitation at 23°C. of a salt-free solution of pH 4.7 into two fractions, one of which is soluble in water at any temperature, and a second one which does not dissolve in water even when heated to 80°C. 2. The proportion of the soluble fraction in gelatin is much greater than of the insoluble one. 3. The insoluble fraction of gelatin does not swell when mixed with water, but it does swell in the presence of acid and alkali which finally dissolve it. 4. Blocks of concentrated gel made by dissolving various mixtures of the soluble and insoluble fractions of gelatin in dilute NaOH swell differently when placed in large volumes of dilute buffer solution pH 4.7 at 5°C. The gel consisting of the insoluble material shows only a trace of swelling, while those containing a mixture of soluble and insoluble swell considerably. The swelling increases rapidly as the proportion of the soluble fraction increases. 5. A 5 per cent gel made up by dissolving the insoluble fraction of gelatin in dilute NaOH loses about 70 per cent of its weight when placed in dilute buffer pH 4.7 at 5°C. A similar gel made up of ordinary gelatin loses only about 20 per cent of its weight under the same conditions. 6. It was not found possible to resynthesize isoelectric gelatin from its components. 7. An insoluble substance similar in many respects to the one obtained by reprecipitation of gelatin is produce on partial hydrolysis of gelatin in dilute hydrochloric acid at 90°C.