The effect of whey protein hydrolyzate average molecular weight on the lactic acid fermentation

Summary The batch fermentation of whey permeate to lactic acid was improved by supplementing the broth with enzyme-hydrolyzed whey protein. Hydrolyzates prepared with endoprotease were more stimulatory to acid production rates than were those prepared with exo/endo protease. The effect of hydrolyzate average molecular weight on acid production is presented. Results show that the hydrolyzate having an average molecular weight of 700 is the most stimulatory to acid production rates.     

Protein partitioning at the isoelectric point: influence of polymer molecular weight and concentration and protein size

We report the partition coefficient, K(p') at the isoelectric point of lysozyme, chymotrypsinogen A, albumin, transferrin, and catalase in 64 different polyethylene(PEG)/ dextran(Dx)/water systems. We study the trends of the partition coefficient with protein type, polymer concentration, and polymer molecular weight. We find that the partition coefficient decreases with increasing tie line length for lysozyme, albumin, transferrin, and catalase for which K(p) is less than 1, but increases for chymotrysinogen for which K(p) is larger than 1. The effect of the tie line length on the partition coefficient is larger for the large proteins than for the small proteins. The partition coefficient decreases with increasing protein molecular weight except for lysozyme suggesting that lysozyme is present as a dimer or a trimer. The partition coefficient decreases with increasing PEG molecular weight, but the magnitude of the increase is larger for the smaller PEG molecular eights and tends to level of at high PEG molecular weight. The partition coefficient increases with increasing dextran (Dx) molecular weight for chymotrypsinogen but decreases for catalase. The partition coefficients of lysozyme, albumin, and transferrin increase with increasing Dx molecular weight from Dx 10(4) to Dx 1.1 x 10(5) and then slightly decrease from Dx 1.1 x 10(5) to Dx 5 x 10(5). The experimental results are analyzed using a statistical thermodynamics model. The experimental results are analyzed using a statistical thermodynamics model. The experiments suggest that protein partitioning at the isoelectric point in aqueous two-phase systems is strongly related to the size of the proteins and polymers. Finally, the impossibility of obtaining data completely independent of polymer concentration is emphasized.     

Isolation and identification of a high molecular weight protein in sow milk

A high molecular weight protein (HMWP) was isolated and purified from sow milk, and some of its biochemical characteristics and biological functions were identified. The origin of HMWP was also investigated. The molecular weight of HMWP was determined to be about 115 000 and 114 800 by SDS-PAGE and gel filtration, respectively. The sequence of 10 amino acids in N-terminal of HMWP was Ala-Leu-Val-Gln-Ser-Cys-Leu-Asn-Leu-Val. The sequence was blasted against GenBank. No protein showed significant similarity with this sequence suggesting the HMWP may be novel. The result of liquid chromatography mass spectrometry (LC-MS) also proved HMWP could be a novel protein. By amino acid assay, HMWP was rich in glutamate (including glutamine), cysteine, glycine, aspartic acid (including asparagines) and proline. The content of hydrophobic amino acids (Ala, Val, Leu, Ile, Met, Phe and Pro) was lower at 18.59% of the total amino acids suggesting HMWP has high solubility in water. Western blots of lectins were used to identify the kinds of carbohydrate residues attached to HMWP qualitatively. The result showed that HMWP was a kind of glycoprotein containing N-acetylneuraminic acid (NeuNAc), mannose (Man) and/or N-acetylglucosamine (GlcNAc). By isoelectric focusing, HMWP pI was found to be 5.1. Compared with milk fat globule membrane protein (MFGMP) isolated from the sow milk in SDS-PAGE, MFGMP did not contain HMWP. HMWP was assumed to be a secretory milk protein. HMWP was not found in bovine, goat, rabbit or human milk in SDS-PAGE gel suggesting HMWP may be unique to sow milk. By Western blot, HMWP could be detected in sow milk, not in sow serum, which suggests it is synthesized and secreted by the mammary gland. HMWP concentrations in sows milk were the lowest in the first day of lactation, rose significantly during lactation 1 to 7 days. The HMWP content of sows milk remained relatively constant ((1.95±0.13) g/l) during lactation 7 to 20 days. HMWP significantly inhibited Escherichia coli in a dose related manner in vitro. Overall, HMWP could be a novel sow milk protein with implications for the mammary gland and the piglet.     

低聚木糖的提取工艺及相对分子质量分布

通过单因素试验和正交试验研究低聚木糖的提取条件,使用凝胶树脂对低聚木糖粗提液进行分离,采用高效液相色谱法测定了低聚木糖的相对分子质量分布。低聚木糖的最佳提取条件:在底物质量浓度为100 g/L的情况下,加酶量1 000 U/g,酶解温度55℃,提取时间为4 h。在此条件下,提取的平均聚合度为3.12,高效液相色谱测定结果发现,低聚木糖主要是由木二糖、木三糖以及4～8个聚合度的糖组成。     

Relationship of amino acid composition and molecular weight of antifreeze glycopeptides to non-colligative freezing point depression

Many polar fishes synthesize a group of eight glycopeptides that exhibit a non-colligative lowering of the freezing point of water. These glycopeptides range in molecular weight between 2600 and 33700. The largest glycopeptides [1–5] lower the freezing point more than the small ones on a weight basis and contain only two amino acids, alanine and threonine, with the disaccharide galactose-N-acetyl-galactosamine attached to threonine. The smaller glycopeptides, 6, 7, and 8, also lower the freezing point and contain proline, which periodically substitutes for alanine. Glycopeptides with similar antifreeze properties isolated from the saffron cod and the Atlantic tomcod contain an additional amino acid, arginine, which substitutes for threonine in glycopeptide 6. In this study we address the question of whether differences in amino acid composition or molecular weight between large and small glycopeptides are responsible for the reduced freezing point depressing capability of the low molecular weight glycopeptides. The results indicate that the degree of amino acid substitutions that occur in glycopeptides 6–8 do not have a significant effect on the unusual freezing point lowering and that the observed decrease in freezing point depression with smaller glycopeptides can be accounted for on the basis of molecular weight.     

功能性低分子量岩藻多糖的研究进展

低分子量岩藻多糖来源于褐藻,是一类含有硫酸基团的多糖,具有多种生物学功能,如抗凝血、抗病毒、抗血栓、抗肿瘤等功能,因此可被广泛地应用于医药、食品等领域。着重介绍了低分子量岩藻多糖的制备及其生物学功能的研究进展。     

低分子肝素对MPTP帕金森小鼠的治疗作用及可能机制

目的：研究1．甲基4-苯基-1,2,3,6-四氢吡啶（1-methy-4-phenyl-1,2,3,6-tetrahy-dropyridine,MPTP）帕金森病（PD）模型中小胶质细胞的激活情况,探讨低分子肝素对MPTP导致的小胶质细胞活化的抑制作用。方法：C57BL随机分成正常对照组、MPTP组、低分子肝素＋MPTP组。MPTP组腹腔注射MPTP（30mg／kgx7d）同时腹部皮下注射生理盐水,低分子肝素＋MPTP组在注射MPTP同时腹部皮下注射低分子肝素（1501U／kg·12hx7d）。各组于末次给药后予行为学测试,7d后免疫组化检测酪氨酸羟化酶（TyrosineHydroxylase,TH）阳性细胞。镀银染色观察小胶质细胞激活情况。结果：MPTP组较低分子肝素＋MPTP组爬竿时间明显延长,并出现更多非随意动作。低分子肝素＋MPTP组黑质部位TH阳性细胞数量高于MPTP组。MPTP组活化的小胶质细胞数量高于低分子肝素＋MPTP组。结论：低分子肝素通过抑制小胶质细胞的激活减少MPTP帕金森小鼠多巴胺能神经元的损伤,提示低分子肝素可能有延缓PD进程的作用。     

The solution molecular weight and shape of the bacterial exopolysaccharides amylovoran and stewartan

Amylovoran, the acidic exopolysaccharide (EPS) of Erwinia amylovora, and stewartan, the capsular EPS of E. stewartii, were characterized by analytical ultracentrifugation and by size exclusion chromatography connected to dual detection of light scattering and mass. The average molecular weights of amylovoran and stewartan were determined as 1.0×10⁶ and 1.7×10⁶ Da, with polydispersity values (M_w/M_n) of 1.5 and 1.4, respectively. Based on the sugar composition and their molecular weight, both exopolysaccharides consist of approximately 1000 repeating units per molecule, this suggests a similar mechanism for chain length determination during biosynthesis of EPS in both organisms.     

低分子肝素的抗炎作用及机制

低分子肝素(low molecular weight heparin, LMWH)除作为抗凝血和抗血栓药在临床上广为应用外,近年来其抗炎活性也颇受重视.LMWH抗炎机制涉及炎症细胞、炎症因子和黏附分子等环节.目前对LMWH的抗炎机制研究还处在初级阶段,但是LMWH独特的性质使其有望成为有效且安全的新型抗炎药物.     

Concerning the use of empirical methods for determining the molecular weight of a small glycopeptide cleaved from bovine rhodopsin

The molecular weight of a glycopeptide cleaved enzymatically from bovine rhodopsin was investigated by a variety of techniques, such as gel filtration, sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence and absence of urea, and gel filtration in 6 guanidinium chloride after reduction and carboxymethylation. Great variation in the estimation of the molecular weight between the several techniques was observed. When compared to an estimate of 9000 based on amino acid analysis and considerations of its carbohydrate content, values ranging between 4500 and 18,000 were obtained by the other methods. Thus, while the molecular weight of the glycopeptide remains unresolved, the resultant ambiguity in evaluating this property may exemplify the limitations inherent in applying empirical methods to the determination of the molecular weight of small glycopeptides.     

Is there a high molecular weight glutamic acid decarboxylase?

Gel-filtration chromatography from crude extracts of mouse brain indicates the presence of a high molecular weight (HMW) (more than 750 kd) and a low molecular weight (LMW) (87.5 Kd) glutamic acid decarboxylase (GAD) when they are concentrated prior chromatography either by precipitation with salts or ethanol. Kinetically both peaks of GAD-activity exhibit an almost identical Km for L-glutamate. Only LMW-GAD appears if the concentration step is carried out by ultrafiltration or if the extract is chromatographied without the concentrating step. HMW-GAD disappears from the chromatographic profiles if ethanolic extracts of GAD are treated with Triton X-100 before the chromatographic procedure. The sodium sulfate precipitation of a previously separated LMW-GAD gives rise to the reappearance of a HMW-GAD peak. Apparently HMW-GAD does not exist as a different molecular entity; indeed it may be an artefactual aggregation of LMW-GAD.     

Separation of both fibrous and globular proteins on the basis of molecular weight using high-performance size exclusion chromatography

A high-performance size exclusion liquid chromatographic system has been used to separate proteins with different shapes solely on the basis of their molecular weights. After the effects of ionic and hydrophobic interactions with the stationary phase have been overcome, protein elution is normally governed by their effective size in solution. Conditions are described under which proteins, with isoelectric points within the normal operating pH range of the columns, are eluted independent of their Stokes' radii. Even fibrous proteins with axial ratios of 50 elute according to their known molecular weights over the range 2000–2,000,000.     

Molecular weight characterization of high molecular weight dextran with multiangle light scattering in on‐line and off‐line mode

This work reports the molecular weight (MW) analysis of high MW dextran using multiangle light scattering (MALS) in both chromatography and automated batch measurement mode. The results show that the chromatographic columns alter the high MW native dextran and cause underestimation of the MW as a consequence. Alternatively, a batch MALS measurement (without columns) provides more accurate MW values. The batch MALS measurement was automated with the incorporation of an automatic sample dilution and injection device. This automation reduces the sample preparation time and minimizes concentration errors introduced by manual sample dilution. To the best of our knowledge, this is the first study using an automated batch MALS in the analysis of high MW dextran. © 2015 Wiley Periodicals, Inc. Biopolymers 103: 387–392, 2015.     

Heme ladder, a direct molecular weight marker for immunoblot analysis

Detection methods for immunoblot analysis are often based on peroxidase conjugates. However, molecular weight markers directly detectable for general use in such systems are not available. Here, we describe the preparation of a direct molecular weight marker consisting of heme-tagged proteins, whose enzymatic activities make them detectable simultaneously with the antigen in peroxidase-based immunoblot systems. The peroxidase activity results from the covalent attachment of heme to selected engineered periplasmic proteins, catalyzed by the cytochrome c maturation system of Escherichia coli. The newly designed heme-tagged proteins were combined with a previously constructed heme-tagged maltose-binding protein and cytochrome c. The resulting heme ladder was shown to be suitable as a protein standard for direct molecular weight estimation in immunoblot analysis due to the peroxidase activity of its constituents. The heme ladder consists of proteins between 12 and 85 kDa and can be produced at low cost. The marker was stable when kept at 4, −20, and −80 °C for >6 months.     

带芒草属物种新型高分子量谷蛋白亚基的鉴定

采用SDSPAGE方法对牧草带芒草属3个种8份材料的高分子量谷蛋白进行了检测和鉴定。结果显示,带芒草物种具有的高分子量谷蛋白亚基与普通小麦中发现的不一样,其迁移率存在较大差异。其中,x型亚基均比Dx2亚基迁移率小或接近,y型亚基均比Dx12亚基迁移率大。8份材料中共发现了4种x型亚基新类型(Tax1,Tax2,Tax3和Tax4),5种y型亚基新类型(Tay1,Tay2,Tay3,Tay4和Tay5)和6种亚基组合类型(Tax1+Tay3,Tax3+Tay2,Tax4+Tay1,Tax1+Tay1,Tax2+Tay5,Tax4+Tay2),该项研究结果揭示了带芒草属植物可能具有与普通小麦类似的高分子量谷蛋白亚基,这些亚基在小麦品质遗传改良中具有潜在的利用价值。     

细菌降解低分子量多环芳烃的研究进展

具有"三致"效应的多环芳烃污染造成了巨大的环境危害,威胁人类健康和生存。目前能够降解低分子量多环芳烃的细菌已有广泛的研究。细菌通过多层次的调控分析和适应性进化提高它们的降解能力。本文基于国内外文献调研,简要总结了生物修复在低分子量多环芳烃降解领域的研究进展。拟通过多层次的调控分析和适应性进化来产生多种分解代谢途径,为生物降解能力强化的未来降解技术提供支撑。     

High recovery of large molecular weight DNA from sorted maize chromosomes

High recovery of large molecular weight DNA from single types of sorted chromosomes is a requirement for the construction of partially digested chromosome-specific libraries. We have developed a simple procedure, based on salting out, for rapidly obtaining large quantities of chromosomal DNA from flow-sorted chromosomes in maize. DNA isolated from sorted frozen chromosomes was of good quality, over 120 kb in size, and suitable for restriction enzyme analysis and construction of phage libraries.     

Optimization of conditions for the production of pullulan and high molecular weight pullulan by Aureobasidium pullulans

Aureobasidium pullulans had a maximum yield coefficient of pullulan (Y _p/s=0.24) with an initial pH of the culture broth of 6.5 in a shake-flask culture. In a batch culture, the maximum pullulan yield coefficient of 0.30 was obtained at the aeration rate of 0.5 vvm. A yeast-like form and mycelial form of cells were found at the culture broth with pH controlled at 4.5 with a maximum yield coefficient of pullulan of 0.27. However, a high portion (35%) of high molecular weight pullulan (M _w>2 000 000) was produced at pH 6.5 with a yeast-like morphology of the cells.     

Reduced point charge models of proteins: assessment based on molecular dynamics simulations

A reduced point charge distribution is used to model Ubiquitin and two complexes, Vps27 UIM-1–Ubiquitin and Barnase–Barstar. It is designed from local extrema in charge density distributions obtained from the Poisson equation applied to smoothed molecular electrostatic potentials. A variant distribution is built by locating point charges on atoms. Various charge fitting conditions are selected, i.e. from either electrostatic Amber99 (Assisted Model Building with Energy Refinement) Coulomb potential or forces, considering reference grid points located within various distances from the protein atoms, with or without separate treatment of main and side chain charges. The program GROMACS (Groningen Machine for Chemical Simulations) is used to generate Amber99SB molecular dynamics (MD) trajectories of the solvated proteins modelled using the various reduced point charge models (RPCMs) so obtained. Point charges that are not located on atoms are considered as virtual sites. Some RPCMs lead to stable MD trajectories. They, however, involve a partial loss in the protein secondary structure and lead to a less-structured solute solvation shell. The model built by fitting charges on Coulomb forces calculated at grid points ranging between 1.4 and 2.0 times the van der Waals radius of the atoms, with a separate treatment of main chain and side chain charges, appears to best approximate all-atom MD trajectories.