Synthesis, in vitro and cellular antioxidant activity evaluation of novel peptides derived from Saccharomyces cerevisiae protein hydrolysate: structure-function relationship : Antioxidant activity and synthetic peptides.

The relationship between structure and function of primary antioxidant peptide, YR-10 (YGKPVAVPAR) was considered by synthesizing three analogues including YHR-10 (YGKHVAVHAR), GA-8 (GKPVAVPA) and PAR-3 (PAR). Antioxidant activity was determined through in vitro and cellular assays. Substitution of Pro with His in the structure of YR-10 led to significant (P < 0.05) higher ABTS radical scavenging and ferric reducing activity. Following in silico simulated gastrointestinal digestion, Tyr and Arg were omitted, respectively, from N and C-terminal positions and resulted in decreasing DPPH, ABTS radical scavenging, and ferric reducing activities. PAR-3 showed the best inhibitory activity on linoleic acid oxidation. Pretreatment of Caco-2 cells with YR-10, YHR-10, and GA-8 (1000 µM) before exposure to H2O2 (160 µM) resulted in 34.10%, 39.66% and 29.159% reduction in malondialdehyde and 53.52%, 17.02% and 24.71% reduction in protein carbonyl levels. The peptide pretreatment reduced catalase level in cells and PAR-3 exhibited the most protective effects on the viability of cells exposed to oxidative stress.

In vitro and in silico studies of novel synthetic ACE-inhibitory peptides derived from Saccharomyces cerevisiae protein hydrolysate.

The structure-function relation of YR-10 (YGKPVAVPAR) was investigated by synthesizing four structural analogs of that including YHR-10 (YGKHVAVHAR), GA-8 (GKPVAVPA), GHA-8 (GKHVAVHA), and PAR-3 (PAR). GA-8 (GKPVAVPA) was synthesized on the basis of simulated enzymatic gastrointestinal digestion performed by bioinformatics tools (expasy-peptide cutter). This study explains the molecular mechanisms for the interaction of synthetic peptides with ACE. The IC50 values of each were 139.554 ±â€¯2.3, 61.91 ±â€¯1.2, 463.230 ±â€¯3.56, 135.135 ±â€¯2.1, 514.024 ±â€¯5.86 µM, respectively. Results indicated that Pro replacement with His in YR-10 and GA-8 increased ACE inhibitory activity respectively, by 55.63% and 70.82%. Removal of Tyr and Arg from respectively N and C terminal positions of YR-10, following in silico simulated gastrointestinal digestion caused the 3.31 fold decrease in ACE inhibitory activity. YHR-10 showed the best docking poses, and GHA-8 exhibited interaction with Zn2+. Lineweaver-Burk plots of most active peptides suggest that they act as noncompetitive inhibitors against ACE.

The Role of Probiotics in Skin Care: Advances, Challenges, and Future Needs.

The skin, being the largest organ in the human body, plays a pivotal role in safeguarding the body against invasive pathogens. Therefore, it is essential to reinforce and protect this vital organ. Current research supports the impact of probiotics on skin health and their ability to alleviate various skin disorders. However, the effectiveness and probable side effects of probiotics in skin care remain a subject of debate, necessitating further investigation and analysis. Hence, this study aims to highlight existing gaps and future needs in the current research on probiotics in skin care and pave the way for future investigations. Therefore, we scrutinized the effects of oral (fermented foods and dietary supplements) and non-oral/topical probiotics on skin care, and the mechanism of probiotics that affect skin health. The results of most studies showed that fermented foods containing probiotics, particularly dairy products, positively impact skin health. The research results regarding the efficacy of probiotic supplements and live strains in treating skin disorders show promising potential. However, safety evaluations are crucial, to identify any potential adverse effects. While research has identified numerous potential mechanisms by which probiotics may influence skin health, a complete understanding of their precise mode of action remains elusive. However, it seems that probiotics can exert their positive effects through the gut-skin and gut-skin-brain axis on the human body. Therefore, following the identification of safe probiotics, additional studies should be carried out to establish optimal dosages, potential side effects, suitable regulatory guidelines, and validation methods.

A microbial inhibition assay in microplates using Bacillus licheniformis for detection of enrofloxacin and sulfamethazine in chicken spiked kidney, liver and muscle tissues.

BACKGROUND: The presence of antimicrobial drugs residues in animal products at levels higher than the maximum residue level (MRL) may have adverse effects on consumer health such as allergic reactions and resistance development. Therefore, it is necessary to monitor animal products for the presence of antimicrobial residues. OBJECTIVES: The aim of this study was to evaluate the detection limit of microbial inhibition assay (MIA) in microplate by using of Bacillus licheniformis as indicator microorganism for two antibiotics, enrofloxacin (ENR) and sulfamethazine (SMT), in broiler chicken's kidney, liver and muscle tissue samples. METHODS: Spiked tissues samples for the two antibiotics were analysed separately by this method. The results of the assay were evaluated by the determination of the absorbance after mean 3.47 h of incubation at 45°C. RESULTS: Results showed that the detection limits of MIA for ENR and SMT in kidney (124.03 and 23.21 µg/kg, respectively) and liver (90.02 and 62.03 µg/kg, respectively) as well as SMT in muscle (46.95 µg/kg) were lower than EU (European Union) - MRL, whereas the detection limit for ENR in muscle was slightly higher than MRL (136.3 µg/kg compared to 100 µg/kg MRL). Furthermore, the MIA in the current study was found to be more sensitive to SMT than ENR (92% and 88% sensitivity rate, respectively). No false-positive was observed in the assay. CONCLUSIONS: Based on the results, the MIA investigated in this study had the potential to detect ENR and SMT residues in broiler chicken kidney, liver and muscle tissues at levels below or close to EU - MRL but offered lower capability for the detection of ENR compared with SMT in kidney and muscle tissue samples.

Improving biomass and carbohydrate production of microalgae in the rotating cultivation system on natural carriers.

Biofilm-based algal technologies have gained popularity due to higher biomass productivity, efficient harvesting, and water-saving over suspended growth systems. A rotating attached system was designed to assess the biofilm-forming capacity of different isolated microalgal strains from the Persian Gulf. Four microalgal strains, including two Chlorella sp., one Picochlorum sp. and one filamentous cyanobacterium Desmonostoc sp. were cultivated on four carriers: jute, cotton, yarn and nylon. The carriers' physicochemical surface characteristics and attachment effects, like contact angle, were investigated. The incorporated biomass and exopolysaccharides (EPS) content in the suspended and biofilm system was calculated and compared. The results showed that the cyanobacterium strain had the biofilm formation capability on both jute and cotton in the attached cultivation system. Under the same culture conditions, the biomass productivity on jute and cotton carriers was significantly higher (4.76 and 3.61 g m- 2 respectively) than the growth in aqueous suspension (1.19 g m- 2 d- 1). The greatest incorporated exopolysaccharides amount was observed on jute (43.62 ± 4.47%) and the lowest amount was obtained from the growth on positive charge yarn (18.62 ± 1.88%). This study showed that in comparison with planktonic growth, the colonization of cyanobacterial cells and subsequent production of extracellular matrix and biofilm formation can lead to increased biomass production.

3D high-precision melt electro written polycaprolactone modified with yeast derived peptides for wound healing.

In this study melt electro written (MEW) scaffolds of poly(ε-caprolactone) PCL are decorated with anti-inflammatory yeast-derived peptide for skin wound healing. Initially, 13 different yeast-derived peptides were screened and analyzed using both in vitro and in vivo assays. The MEW scaffolds are functionalized with the selected peptide VLSTSFPPW (VW-9) with the highest activity in reducing pro-inflammatory cytokines and stimulating fibroblast proliferation, migration, and collagen production. The peptide was conjugated to the MEW scaffolds using carbodiimide (CDI) and thiol chemistry, with and without plasma treatment, as well as by directly mixing the peptide with the polymer before printing. The MEW scaffolds modified using CDI and thiol chemistry with plasma treatment showed improved fibroblast and macrophage penetration and adhesion, as well as increased cell proliferation and superior anti-inflammatory properties, compared to the other groups. When applied to full-thickness excisional wounds in rats, the peptide-modified MEW scaffold significantly enhanced the healing process compared to controls (p < 0.05). This study provides proof of concept for using yeast-derived peptides to functionalize biomaterials for skin wound healing.

Characterization, antioxidant and anticoagulant properties of exopolysaccharide from marine microalgae.

The sulfated exopolysaccharide extracted from marine microalgae attracted considerable attention from both the nutraceutical and pharmaceutical industries. In the present study biomass of five marine microalgae were screened to find strains with high capacity for the production of sulfated exopolysaccharides. The anticoagulant and antioxidant activities of extracted sulfated polysaccharides were evaluated using activated partial thromboplastin time (aPTT), prothrombin time (PT), DPPH and ABTS assays, respectively. The sulfated polysaccharides extracted from Picochlorum sp. showed a strong DPPH scavenging effect with 85% antioxidant activity. The sulfated polysaccharides of Chlorella sorokiniana, Chlorella sp. (L2) and Chlorella sp. (D1) scavenged more than 90% of the ABTS radicals. However, the sulfated polysaccharide extracted from Chlorella sorokiniana, and Chlorella sp. (N4) showed anticoagulant properties. The dual anticoagulant-antioxidant activities in Chlorella sorokiniana could be explained by the combination of various factors including sulfate content and their binding site, monosaccharide residue and glycoside bond which are involved in the polysaccharide's bioactivity.

High-level expression of lipase in Escherichia coli and recovery of active recombinant enzyme through in vitro refolding.

Microbial lipases are widely used for biotechnological applications. In this study we have cloned and sequenced the lipase and lipase specific foldase genes of a Pseudomonas sp., which was isolated from Southern Iran. The lipase was composed of 936 bp which encoded 311 amino acids and the lipase specific foldase gene consisted of 1008 bp which encoded 336 amino acids. The low amount of recombinant lipase was expressed as an active enzyme in Escherichia coli harboring a plasmid with the clustered lipase and lipase specific foldase genes. To increase the enzyme expression level, the lipase and lipase specific foldase genes subcloned into two separate expression vectors. The lipase was expressed as inactive inclusion bodies under the control of the strong T7 promoter. Inclusion bodies were dissolved in 8M urea and 1mM dithiothreitol (DTT) and purified using Ni-nitrilotriacetic acid column. Subsequently, purified lipase diluted in 20mM phosphate buffer (pH 7) containing the lipase specific foldase which was expressed in another clone of E. coli. In the presence of foldase, it was possible to achieve active lipase with a specific activity of up to 240 IU/mg using a simple refolding procedure. Moreover, the effect of different concentrations of various additives was investigated on the refolding of denatured lipase. The best yield of 70 IU/ml with the specific activity of 3000 IU/mg were obtained after incubation of denatured enzyme in a refolding buffer containing lipase specific foldase (0.005 mg/ml), 1M NaCl and 10% glycerol at 4 degrees C.

Effects of temperature treatment on corn cob composting and reducing of composting time: a comparative study.

This study focuses on the effect of temperature on the compost quality and the processing time. Two temperature, namely 46°C (high temperature) and 30°C (low temperature) were considered for the composting trials. A mixture of 60% ground corn cob, 30% dried sports grass and 10% bio-activator was used as composting material. The composting process lasted for 120 days. The maturity indices at the end of composting indicate that, for the high-temperature treatment, the compost show an improved quality as compared to the low-temperature treatment. The maturity indices, namely carbon to nitrogen ratio, water extractable organic carbon, ratio of cation exchange capacity to total organic carbon and germination index, determined for the high temperature treatment are: 13.9, 1.48 mg mL⁻¹, 2.21, and 86%, respectively. On the basis of the compost characteristics, the processing time for the high- and low-temperature treatments was 75 days and 105 days, respectively. It can be concluded that treatment at the high-temperature level considered in this study presents a more suitable option for improvement of composting.

Comparison of Different Nisin Separation and Concentration Methods: Industrial and Cost-Effective Perspectives.

Separation and concentration of biotechnological products are the most important steps of purification stage in downstream processing. In the present study, three nisin separation and concentration methods including salting out by ammonium sulfate, solvent extraction (at 20 °C, 4 °C, and - 10 °C) and direct chromatography of culture medium were compared with each other. According to our results, nisin precipitation by ammonium sulfate at 40% saturation was the most efficient method (yield = 90.04%, purification fold = 168.80). Low yield and fold purification values were obtained by solvent extraction with chloroform (yield = 24.23%, fold purification = 37.43 at - 10 °C) and direct cation exchange chromatography of culture medium (yield = 20.00%, fold purification = 1.80). Also, performing purification steps in low pH values and acidic conditions (pH = 3.0) is essential for efficient nisin purification.

Screening of carbon and nitrogen sources using mixture analysis designs for carotenoid production by Blakeslea trispora.

The production of many secondary metabolites such as carotenoids is influenced by the type of carbon and nitrogen sources and C:N ratio applied in culture medium. The present study discusses the role of C:N ratio and screening of carbon and nitrogen sources using mixture analysis design in carotenoids production by Blakeslea trispora. The C:N ratios of 20, 40, and 60 with six nitrogen sources were evaluated. Results indicated that limitation of nitrogen source (C:N of 60) could improve carotenoids production. Six nitrogen and carbon sources were then screened using mixture analysis design. The most effective nitrogen and carbon sources were soybean powder and glucose, respectively. The productivity of carotenoids (983.8 ± 31.5 mg/L) based on consumed nitrogen and carbon source was 189.10 mg/g soybean powder and 19.66 mg/g glucose. Mixture analysis design indicated single carbon and nitrogen source were more effective than a mixture of sources for carotenoids production.

Production of antioxidant and ACE-inhibitory peptides from Kluyveromyces marxianus protein hydrolysates: Purification and molecular docking.

Kluyveromyces marxianus protein hydrolysates were prepared by two different sonicated-enzymatic (trypsin and chymotrypsin) hydrolysis treatments to obtain antioxidant and ACE-inhibitory peptides. Trypsin and chymotrypsin hydrolysates obtained by 5 h, exhibited the highest antioxidant and ACE-inhibitory activities. After fractionation using ultrafiltration and reverse phase high performance liquid chromatography (RP-HPLC) techniques, two new peptides were identified. One fragment (LL-9, MW = 1180 Da) with the amino acid sequence of Leu-Pro-Glu-Ser-Val-His-Leu-Asp-Lys showed significant ACE inhibitory activity (IC50 = 22.88 µM) while another peptide fragment (VL-9, MW = 1118 Da) with the amino acid sequence of Val-Leu-Ser-Thr-Ser-Phe-Pro-Pro-Lys showed the highest antioxidant and ACE inhibitory properties (IC50 = 15.20 µM, 5568 µM TE/mg protein). The molecular docking studies revealed that the ACE inhibitory activities of VL-9 is due to interaction with the S2 (His513, His353, Glu281) and S'1 (Glu162) pockets of ACE and LL-9 can fit perfectly into the S1 (Thr345) and S2 (Tyr520, Lys511, Gln281) pockets of ACE.

Optimization of Non-Nutritional Factors for a Cost-Effective Enhancement of Nisin Production Using Orthogonal Array Method.

In order to increase nisin production in a cost-effective manner, non-nutritional factors as well as nutritional parameters must be optimized. In this study, optimization of the most important non-nutritional factors for nisin production using orthogonal array method was performed. Optimization of temperature, agitation, age and size of inoculum, medium initial pH value and flask volume/medium volume ratio in de Man, Rogosa and Sharpe (MRS) medium in batch fermentation was accomplished. Nisin was produced by Lactococcus lactis subsp. lactis PTCC 1336 and measured by bioassay method using Micrococcus luteus PTCC 1169 as the nisin-sensitive strain. The optimum levels of non-nutritional factors for maximum nisin production and productivity were obtained as: flask volume/medium volume ratio: 5.00, medium initial pH value: 8.00, inoculum size: 1%, inoculum age: 24 h old (A = 1.7), agitation: 100 rpm and temperature: 27 °C. Under the optimized conditions, maximum nisin production and maximum nisin productivity were 599.70 IU/mL and 37.48 IU/mL/h, respectively.