Biomimetic assembly of proteins into microcapsules on oil-in-water droplets with structural reinforcement via biomolecular-recognition-based cross-linking of surface peptides

By mimicking the stabilization of bacterial membranes with S-layer proteins, a novel process to fabricate highly stable protein microcapsules is introduced. In this strategy, engineered collagen peptides with site-specific biotinylation are assembled into microcapsules on the oil-in-water droplets, and the resulting microcapsules are reinforced by biomolecular-recognition-based cross-linking with the protein. Furthermore the microcapsules are shown to be versatile scaffolds for developing functionalized hierarchical colloidosomes for important biotechnological applications.     

Engineering Functional Membrane–Membrane Interfaces by InterSpy

Engineering synthetic interfaces between membranes has potential applications in designing non-native cellular communication pathways and creating synthetic tissues. Here, InterSpy is introduced as a synthetic biology tool consisting of a heterodimeric protein engineered to form and maintain membrane–membrane interfaces between apposing synthetic as well as cell membranes through the SpyTag/SpyCatcher interaction. The inclusion of split fluorescent protein fragments in InterSpy allows tracking of the formation of a membrane–membrane interface and reconstitution of functional fluorescent protein in the space between apposing membranes. First, InterSpy is demonstrated by testing split protein designs using a mammalian cell-free expression (CFE) system. By utilizing co-translational helix insertion, cell-free synthesized InterSpy fragments are incorporated into the membrane of liposomes and supported lipid bilayers with the desired topology. Functional reconstitution of split fluorescent protein between the membranes is strictly dependent on SpyTag/SpyCatcher. Finally, InterSpy is demonstrated in mammalian cells by detecting fluorescence reconstitution of split protein at the membrane–membrane interface between two cells each expressing a component of InterSpy. InterSpy demonstrates the power of CFE systems in the functional reconstitution of synthetic membrane interfaces via proximity-inducing proteins. This technology may also prove useful where cell-cell contacts and communication are recreated in a controlled manner using minimal components.     

低灰分血浆蛋白粉制备工艺优化

为有效利用动物血液,优化血浆蛋白粉的制备工艺,本文从分离、浓缩与喷雾干燥工艺出发,探讨了不同的离心转速和离心时间对血浆蛋白总浓度、血红蛋白残留量及收集率的影响,不同的超滤压力、p H对血浆蛋白粉水分、粗蛋白及灰分含量的影响,不同的进口温度和进料速率对血浆蛋白粉水分含量、粗蛋白、提取率及氮溶解指数的影响。结果表明:最佳离心条件为离心转速5000 r/min,离心时间10 min;最佳超滤条件为超滤压力0.6 MPa,血浆液p H为9;最佳喷雾干燥工艺为进口温度150℃,进料速率0.178 m L/s。经本工艺优化,血浆蛋白粉的灰分降至6%左右。本研究结果可为低灰分血浆蛋白粉生产与制备工艺的改进提供基础数据和参考。      

Physico-chemical characteristics and non-starch polysaccharide contents of Indica and Japonica brown rice and their malts

Brown rice malt from Indica and Japonica type rice were prepared and their nutrient composition as well as non-starch polysaccharide (NSP) contents and also some of the physico-chemical characteristics were determined. The activity of α- and β-amylases in the ungerminated (native) rice was negligible but increased considerably on germination. Malting altered the chemical composition of both Indica and Japonica rice to a small extent but caused noticeable changes in the pasting characteristics. The free sugars and water-soluble non-starch polysaccharide (NSP) contents of the malt samples were considerably higher than the native samples. The carbohydrate elution profile of native and malted rice on Sepharose CL-2B revealed partial degradation of starch. However, the electrophoresis patterns of the native and malt samples were comparable but exhibited decrease in their band intensity. The study revealed that, controlled germination or malting causes considerable changes in the physico-chemical and bio-chemical properties of both Indica and Japonica rice.     

Heat stability of reconstituted, protein-standardized skim milk powders

We determined the effects of standardization material, protein content, and pH on the heat stability of reconstituted milk made from low-heat (LH) and medium-heat (MH) nonfat dry milk (NDM). Low-heat and MH NDM were standardized downward from 35.5% to 34, 32, and 30% protein by adding either edible lactose powder (ELP) or permeate powder (PP) from skim milk ultrafiltration. These powders were called standardized skim milk powders (SSMP). The LH and MH NDM and SSMP were reconstituted to 9% total solids. Furthermore, subsamples of reconstituted NDM and SSMP samples were set aside to measure heat stability at native (unadjusted) pH, and the rest were adjusted to pH 6.3 to 7.0. Heat stability is defined as heat coagulation time at 140°C of the reconstituted LH or MH NDM and SSMP samples. The entire experiment was replicated 3 times at unadjusted pH values and 2 times at adjusted pH values. At an unadjusted pH, powder type, standardization material, and protein content influenced the heat stability of the samples. Heat stability for reconstituted LH NDM and SSMP was higher than reconstituted MH NDM and SSMP. Generally, decreased heat stability was observed in reconstituted LH or MH SSMP as protein content was decreased by standardization. However, adding ELP to MH SSMP did not significantly change its heat stability. When pH was adjusted to values between 6.3 and 7.0, powder type, standardization material, and pH had a significant effect on heat stability, whereas protein content did not. Maximum heat stability was noted at pH 6.7 for both reconstituted LH NDM and SSMP samples, and at pH 6.6 for both reconstituted MH NDM and SSMP samples. Furthermore, for samples with adjusted pH, higher heat stability was observed for reconstituted LH SSMP containing PP compared with reconstituted milk from LH SSMP containing ELP. However, no statistical difference was observed in the heat stability of reconstituted milk from MH NDM and MH SSMP samples. We conclude that powder type (LH or MH) and effect of standardization material (ELP or PP) can help explain differences in heat stability. The difference in the heat stability of powder type may be associated with the difference in the pH of maximum heat stability and compositional differences in the standardization material (ELP or PP).     

Assessment of protein changes in farmed giant catfish (Pangasianodon gigas) muscles during refrigerated storage

Farmed giant catfish (Pangasianodon gigas) muscles (dorsal and ventral sites) were stored in a refrigerator (at 4 °C) for 14 days to determine the effect of refrigerated storage on biochemical and physical changes. The analyses were carried out at 0, 2, 4, 7, 10 and 14 days of storage. At day 14, Ca²⁺ ‐ATPase activity markedly decreased when compared to its value at day 1 (>90%), while a small decrease was observed for surface hydrophobicity and reactive sulfhydryls content. Total volatile basic nitrogen and trichloroacetic‐soluble peptide content gradually increased when the storage period was extended. The myosin heavy chain decreased slightly on SDS‐PAGE for both meat cuts with increased storage time. Expressible drip and cooking loss were highest during the first day of storage and slightly decreased with storage time. Instrumental hardness was significantly higher in the ventral compared to the dorsal muscle, while the toughness was the highest at the second day of storage. The muscle bundles with scanning electron microscopy were less attached, resulting in the observed big gaps over increasing storage time. Results indicated that changes of proteins have detrimental effects on the quality attributes of farmed giant catfish muscles during refrigerated storage, particularly physical and biochemical properties.     

Extraction and characterisation of protein fractions from five insect species

Tenebrio molitor, Zophobas morio, Alphitobius diaperinus, Acheta domesticus and Blaptica dubia were evaluated for their potential as a future protein source. Crude protein content ranged from 19% to 22% (Dumas analysis). Essential amino acid levels in all insect species were comparable with soybean proteins, but lower than for casein. After aqueous extraction, next to a fat fraction, a supernatant, pellet, and residue were obtained, containing 17–23%, 33–39%, 31–47% of total protein, respectively. At 3% (w/v), supernatant fractions did not form stable foams and gels at pH 3, 5, 7, and 10, except for gelation for A. domesticus at pH 7. At 30% w/v, gels at pH 7 and pH 10 were formed, but not at pH 3 and pH 5. In conclusion, the insect species studied have potential to be used in foods due to: (1) absolute protein levels; (2) protein quality; (3) ability to form gels.     

Modulation of the antioxidant/pro-oxidant balance,cytotoxicity and antiviral actions of grape seed extracts

Grape seed extracts (GSEs) were investigated in yeast cells harbouring defects in their antioxidant system (regarding the cellular growth and growth recovery from H₂O₂ insult). GSEs antioxidant activity was detected in wild-type and mutant strains Δcta1, Δgsh1 and Δoye2glr1, while pro-oxidant activity in Δsod1 cells was seen. Assessment of proliferation of prostate cancer PC3 and HBV-replicating HepG2 2.2.15 cells treated with GSEs has shown higher cytotoxicity of red grape seed extract (RW) than white grape seed extract (WW) subjective to dose and period of administration. No antiviral effect was detected by measuring the secreted virion particles in HepG2 2.2.15 cells treated with GSEs. The GSEs play a dual antioxidant/pro-oxidant role in vivo according with the cellular antioxidant system deficiencies and exhibit cytotoxic properties in PC3 and HepG2 2.2.15 cell lines, but no antiviral action against HBV.     

Influence of fortification with sodium–calcium caseinate and whey protein concentrate on microbiological,textural and sensory properties of set‐type yoghurt

The effect of fortification of yoghurt with sodium–calcium caseinate (SCC) and whey protein concentrate (WPC) on some properties of set‐type yoghurt were investigated. The addition of WPC enhanced the viability of Lactobacillus delbrueckii subsp. bulgaricus more than SCC. The highest firmness values were obtained from SCC‐fortified yoghurts, whereas yoghurts fortified with WPC had the highest water‐holding capacity during storage. The yoghurts fortified with 4% w/w SCC or 4% w/w WPC had the highest viscosity. Yoghurts fortified with 2% w/w SMP, SCC or WPC showed similar taste and overall acceptability scores; however, samples containing 4% w/w SCC or 4% w/w WPC had the lowest scores.     

Partial biochemical characterization of tetrazolium red‐reactive Lactobacillus plantarum mutants

One thousand colonies derived from Lactobacillus plantarum ATCC 8014 cells that survived 34–43 × 10³ ergs cm^‐2 ultraviolet irradiation were screened on media containing tetrazolium red to detect fermentative mutants. Fermentation end‐products formed from pyruvate, glucose, or lactose catabolism were determined. All 37 stable tetrazolium red‐reactive mutants had increased pyruvate utilization compared to the wild‐type strain. Only two did not produce lactate from exogenous pyruvate. When glucose or lactose were substrates, these two mutants and six other representative mutants produced lactate at levels similar to the parent strain. Although the average lactate and acetoin production from pyruvate by the mutants and wild‐type strains were similar, 25% of the mutants had increased acetoin production.