Collagenase Activity Research Articles

Solar radiation leads to increased collagenase gene expression (MMP1) in HERDA (Hereditary equine regional dermal asthenia) affected horses and may explain the typical dorsal distribution of their lesions.

Hereditary equine regional dermal asthenia (HERDA) is a genetic disease that alters collagen biosynthesis. Affected horses exhibit fragile, hyperextensible skin, especially over the dorsal region. Although ultraviolet (UV) radiation seems to contribute to the regional distribution of lesions and worsening of clinical signs, the molecular mechanisms involved are largely unknown. To evaluate the effect of solar radiation on matrix metalloproteinase MMP1, MMP8 and MMP13 gene expression in the dorsal and ventral skin of HERDA-affected and HERDA-unaffected horses [wild-type (WT) horses]. Six HERDA-affected and six unaffected Quarter horses (WT) were paired according to age, sex and coat colour. Horses were submitted to 30 day sunlight restriction, followed by 15 day sunlight exposure. Dorsal and ventral skin biopsies were obtained at six sampling times over 45 days. The expression of MMP1, MMP8 and MMP13 genes was measured by quantitative PCR. Although solar radiation modulated MMP1, MMP8 and MMP13 expression, the effects were more pronounced on MMP1. Sun exposure for three days significantly upregulated MMP1 in the dorsal region when compared to the ventral skin in both unaffected and HERDA-affected horses. This study shows that solar irradiation leads to upregulation of skin collagenase genes particularly MMP1 in the dorsal, sun-exposed skin of horses. Furthermore, this was more marked in HERDA-affected horses. The increased activity of collagenases on the disorganised collagen present in HERDA affected horses would explain why UV radiation leads to deterioration of clinical signs in affected individuals.

A two-phase and long-lasting multi-antibacterial coating enables titanium biomaterials to prevent implants-related infections.

In clinical work, the main challenges for titanium (Ti) implantation are bacterial infection and aseptic loosening, which severely affect the survival rate of implants. The first 4 weeks post-operation is the infection peak phase of implants. Inhibiting implant infection caused by bacteria adhesion and proliferation during the early phase as well as promoting subsequent osteointegration is essential for implant success. Herein, we constructed a quaternary ammonium carboxymethyl chitosan (QCMC), collagen (COL Ⅰ) and hydroxyapatite (HAP) multilayers coating on Ti substrates via a modified layer-by-layer (LBL) technique and polymerization of dopamine. The QCMC/COL/HAP coating exhibited a multi-antibacterial property with a two-phase function: (1) At the first 4 weeks post-operation, the covalently bonded QCMC could be slowly degraded and demonstrated both contact-killing and release-killing properties during the infection peak phase; (2) At the second phase, osteogenesis and osseointegration-promotion capabilities were enhanced by HAP under the effective control of infection. The multifilm coating was degraded for more than 45 days under the action of collagenase Ⅰ, and displayed good biocompatibility in vivo and in vitro. Most importantly, the coating exhibited a long-lasting antibacterial activity for more than 3 months, against the main pathogenic bacteria of peri-implant infections. Both in vitro studies and in vivo animal models revealed a desirable osteogenic differentiation capacity of Ti-CCH. Therefore, our study reports a two-phase, long-lasting multi-antibacterial coating on Ti-CCH and indicates potential applications of the modified LBL strategy in orthopaedic fields, which is enlightening for developing practical implant and scaffold materials.

Cyclic carbonate: A green multifunctional agent for sustainable leather manufacture

“Go Green” manoeuvre is gaining prominence due to environmental sustainability, where chemical consumption needs to be subjugated. In order to alleviate this issue, industries have adopted the use of multifunctional chemicals as an alternative to mono-functional agents. The global interest in cyclic carbonate is emerging because of its solvent-cum-reagent characteristic. In this work, we have attempted to use “Propylene Carbonate (PC)” as a green multifunctional agent in the different unit processes of leather making. The antimicrobial activity of PC against gram-negative (Bacillus subtilis, Staphylococcus aureus), gram-positive (Escherichia coli, Klebsiella pneumonia) bacterial strains, and mixed culture prepared from putrefied skin extract enables to use of PC in the preservation process as an alternative to common salt (NaCl). The goatskins treated with 15% PC can be stored for up to 2 months without any putrefaction. Further, the developed method reduces the 85% of TDS load from the soaking process and also completely eliminates the generation of contaminated salt. Spectroscopic investigation and molecular dynamic simulation studies clearly indicate that the structural stability of collagen is not affected in the presence of PC. However, it inhibits the activity of collagenase on collagen. The use of PC as an alkali neutralizing agent in the deliming process completely avoids the generation of toxic ammonium gas. The possibility of using PC as an antifungal agent in the chrome tanning process has been attempted. The wet-blue leather treated with 3% PC has been preserved for 30 days without any fungal attack. Hence, the application of PC in various unit processes has a great potential in reducing chemical consumption thereby leading to sustainable leather manufacturing.

Coriander (Coriandrum sativum L.) essential oil and oil-loaded nano-formulations as an anti-aging potentiality via TGF\u03b2/SMAD pathway

Aging has become a concern for many people, especially women. Given that high-quality anti-aging products are of high cost; it has imperative to search for other economical sources. Essential oils are frequently used in cosmetics products due to a wide range of biological activities as well as their pleasant odor. The current study aimed to investigate the biochemical effect of the cosmetic potential of selected Apiaceous essential oils, traditionally used for skincare, by evaluating their anti-wrinkle activity. It is worth noting that, coriander essential oil showed the highest collagenase, elastase, tyrosinase, and hyaluronidase inhibitory activities compared to other Apiaceous oils (fennel, anise, and cumin). GC–MS proved that coriander essential oil showed a very high level of oxygenated monoterpenes, with linalool (81.29%) as the most abundant constituent. Intriguingly, coriander oil cream and Coriander Essential Oil-loaded Lipid Nanoparticles (CEOLNs) formulations attenuated in vivo UV-induced skin photoaging that was manifested by significantly decreased MDA, COX-2, PGE-2, MMP-1, JNK, and AP-1 levels. Moreover, these pharmaceutical dosage forms significantly increased skin collagen content compared to UV-injured group. Also, coriander essential oil significantly increased TGFβ, TGFβII, and SMAD3 protein expression levels compared to UV-injured group. In conclusion, the pharmaceutical dosage forms of coriander oil possess anti-wrinkle activities that could have an auspicious role in amending extrinsic aging.

An Investigation of the Altered Textural Property in Woody Breast Myopathy Using an Integrative Omics Approach.

Woody breast (WB) is a myopathy observed in broiler Pectoralis major (PM) characterized by its tough and rubbery texture with greater level of calcium content. The objective of this study was to investigate the functionality/integrity of WB sarcoplasmic reticulum (SR), which may contribute to the elevated calcium content observed in WB and other factors that may influence WB texture. Fourteen Ross line broiler PM [7 severe WB and 7 normal (N)] were selected, packaged, and frozen at −20°C at 8 h postmortem from a commercial processing plant. Samples were used to measure pH, sarcomere length, proteolysis, calpain activity, collagenase activity, collagen content, collagen crosslinks density, and connective tissue peak transitional temperature. Exudate was also collected from each sample to evaluate free calcium concentration. The SR fraction of the samples was separated and utilized for proteomic and lipidomic analysis. The WB PM had a higher pH, shorter sarcomeres, lower % of intact troponin-T, more autolyzed μ/m calpain, more activated collagenase, greater collagen content, greater mature collagen crosslinks density, and higher connective tissue peak transitional temperature than the N PM (p ≤ 0.05). Exudate from WB PM had higher levels of free calcium than those from N PM (p < 0.05). Proteomics data revealed an upregulation of calcium transport proteins and a downregulation of proteins responsible for calcium release (p < 0.05) in WB SR. Interestingly, there was an upregulation of phospholipase A2 (PLA2), and cholinesterase exhibited a 7.6-fold increase in WB SR (p < 0.01). Lipidomics data revealed WB SR had less relative % of phosphatidylcholine (PC) and more lysophosphatidylcholine (LPC; p < 0.05). The results indicated that upregulation of calcium transport proteins and downregulation of calcium-release proteins in WB SR may be the muscle’s attempt to regulate this proposed excessive signaling of calcium release due to multiple factors, such as upregulation of PLA2 resulting in PC hydrolysis and presence of cholinesterase inhibitors in the system prolonging action potential. In addition, the textural abnormality of WB may be the combined effects of shorter sarcomere length and more collagen with greater crosslink density being deposited in the broiler PM.

Microemulsion of essential oils from citrus peels and leaves with anti-aging, whitening, and irritation reducing capacity

Although citrus essential oil is frequently used in cosmetics as fragrance, the biological activities related to skin benefits have been rarely investigated. This study aimed to investigate the anti-skin aging and anti-melanogenic effects of citrus oils and develop microemulsion system to reduce the irritation capacity of citrus oils. Oils from leaves and fruit peels of Citrus aurantifolia (Christm.) Swing and Citrus reticulata Blanco cv. Shogun were from hydro-distillation. Their chemical composition was analyzed using gas chromatography/mass spectrometry. Antioxidant activities were determined in terms of free radical scavenging activities, ferric reducing antioxidant power, and lipid peroxidation inhibition. Anti-aging and anti-melanogenic activities were investigated using related enzyme activity analysis. Oils with the most potent biological activities were selected for further incorporation into microemulsion. The irritation capacity of native and citrus microemulsion were investigated in human volunteers. Citrus oils comprised a range of chemicals, with limonene being a key component. All citrus oils demonstrated powerful antioxidative activities. C. reticulata dried peel oil significantly inhibited collagenase and tyrosinase activities, resulting in promising anti-aging and whitening properties. C. reticulata oil derived from dried peels (CR3) had a higher yield and more potent activity than the oils taken from fresh leaves and peels. Therefore, CR3 was developed into a microemulsion in which the internal droplet size was 266.5 ± 14.1 nm and the narrowest PDI of 0.396 ± 0.068. Microemulsion could significantly reduce the skin irritation property of CR3 in human volunteers. In conclusion, CR3 microemulsion was suggested as effective and safe for further cosmeceutical applications.

Development of Ultrasound-Assisted Extraction to Produce Skin-Whitening and Anti-Wrinkle Substances from Safflower Seed.

In this study, ultrasound-assisted extraction (UAE) was applied to extract bioactive substances with skin-whitening, anti-wrinkle, and antioxidant effects from safflower seeds, and the extraction conditions were optimized by a central composite design. The independent variables, including extraction time (5.0~55.0 min), extraction temperature (26.0~94.0 °C), and ethanol concentration (0.0~100%), were optimized to increase tyrosinase activity inhibitory (TAI), collagenase activity inhibitory (CAI), and radical scavenging activity (RSA), which are indicators of skin-whitening, anti-wrinkle, and antioxidant effects. An extraction time of 26.4 min, extraction temperature of 52.1 °C, and ethanol concentration of 50.7% were found to be optimum conditions of UAE, under which TAI, CAI, and RSA were 53.3%, 91.5%, and 27.7%, respectively. The extract produced by UAE was analyzed by LC-MS/MS, and maleic acid and levulinic acid were identified as the main substances. Therefore, UAE is evaluated as an effective process to extract skin-whitening, anti-wrinkle, and antioxidant substances from safflower seeds at lower temperatures and shorter extraction times compared to the conventional extraction methods. Overall, safflower seeds extract can be used as a material for value-added cosmetics, including maleic acid and levulinic acid, which have bioactive functions.

Molecular Docking, Tyrosinase, Collagenase, and Elastase Inhibition Activities of Argan By-Products

The argan tree (Argania spinosa (L.) Skeels) is one of the most important floristic resources in Morocco. This Moroccan endemic tree is known for its numerous therapeutic and medicinal uses. In addition to some medicinal and cosmetic uses, argan fruit pulp and press cake are traditionally used by the Berber population for heating and feeding livestock. Molecular docking is an in silico approach that predicts the interaction between a ligand and a protein. This approach is mainly used in chemistry and pharmacology of natural products as a prediction tool with the purpose of selecting plant extracts or fractions for in vitro tests. The aim of this research is to study the evaluation of potential tyrosinase, collagenase, and elastase inhibitory activities of argan fruit press-cake and pulp extracts. Extracts were evaluated for their total phenolic content (TPC), and the major polyphenols of both press-cake and pulp extracts were submitted to molecular docking in order to determine the mechanisms of action of these compounds. Obtained results revealed that fruit pulp had the strongest dermocosmetic activities, as well as the highest TPC, with values above 55 mg gallic-acid equivalent per gram of dry matter (mgeq AG/gDM). Moreover, those results were positively correlated with the docking findings, suggesting that the pulp lead compounds have higher affinity with tyrosinase, collagenase, and elastase action sites. The results here presented are very promising and open new perspectives for the exploitation of argan-tree by-products as cosmetic agents towards the development of new anti-aging products.

First report of collagenase production by Trichosporon sp. strain isolated from pollen of Amazonian bee (Melipona seminigra seminigra)

Trichosporon yeasts are widely employed to produce lipids, lipases, and aspartic peptidases, but there are no previous studies on collagenase production. This work aimed to select the best collagenase producing Amazonian Trichosporon strains. Moreover, a 23-full factorial design (FFD) and a 22-central composite design combined with Response Surface Methodology were applied to optimize production and find the best conditions for hydrolysis of type I bovine collagen. Most of the studied strains had some collagenolytic activity, but the selected one achieved the highest value (44.02 U) and a biomass concentration of 2.31 g/L. The best collagenase production conditions were 160 rpm of agitation, pH 5.5 and a substrate concentration of 4.0 g/L. The former experimental design showed that substrate concentration was the only statistically significant factor on both biomass concentration and collagenase activity, while the latter showed simultaneous effects of substrate concentration and pH on collagenolytic activity, which peaked at pH 5.5–6.4 and substrate concentration of 3.0–3.4 g/L. An additional 2³-FFD was finally used to optimize the conditions collagen hydrolysis, and pH 6, 25 °C and a substrate concentration of 7.5 (g/L) ensured the highest hydrolysis degree. This study is the first that describes optimized conditions of collagenase production by Trichosporon strains.

Medicinal significance of drug-like compounds derived from Anethum sowa L. seed oil and in-silico predictive investigation of cancer therapy potential

Anethum sowa L. is an annual sweet-smelling herb recognized for gastronomic besides therapeutic usage since primeval times. Chemical configuration and the medicinal utility of seed essential oil of this herb grown in South Karnataka is limited; furthermore, the volatile oil and extract has not been deliberated or compared for its out-turn on Human Colorectal Carcinoma cell line -HCT -116. The current study was envisioned to recognize the components present in the seed oil of Anethum sowa L. examined by GC-MS. Cyclohexanone,2-methyl-5-(1-methylethenyl)-, trans-(41.66%),cyclodecene,1-methyl-(28.15%),(1R,2R)-1,2-bis(prop-1-en-2-yl)cyclobutene(9.80%),(5S)-6,6-Dimethyl-2-azaspiro[4.4]nona-1-ene (1.05%), Benzene, 1-methoxy-4-methyl-2-[[[2-(4-nitrophenyl)hydrazine}carbonyl]amino]-(0.74%),L-Limonene(0.26%) were the new compounds found and reported.2,2-diphenyl-1-picryl-hydrazyl-hydrate test of extract exhibited a half-maximal inhibitory concentration of 640.129 μg/ml. The essential oil of seed showed effective antimicrobial activity against Klebsiella pneumoniae strain (MTCC 3384) &amp; Streptococcus mutants’ strain (MTCC 497) with a MIC of 20% and 10% respectively. Anticancer potential against HCT 116 exhibited that the seed oil and seed methanolic extract suppressed the cell progress in a concentration hooked on method, where seed oil with an IC50 99.38μg/ml was more active when compared with the extract. Seed oil also exhibited potent ant collagenase activity of 56.027% whereas extract showed only 34.312%. In silico docking was performed with the selected compounds of high concentration present in the seed oil with Adenomatous Polyposis Coli of Homo sapiens and MAI-150 peptidomimetic inhibitor as control. D-Limonene showed the maximum binding affinity with APC compared to the rest of the other compounds present in the Seed oil, which in turn confers the anticancer us property for the Seed oil. Keywords: Anticancer, Antimicrobial, Antioxidant, Anticollagense, Colorectal cancer, D-Limonene.

Collagenase Activity of Vaginal Microbes Associated with Preterm Labour

Collagens in the cervix (type I) and chorioamniotic membranes (type I, IV) play important biomechanical roles in maintaining healthy pregnancy and undergo significant modulation during natural labour progression. However, timing is key: inappropriate remodeling of collagens can trigger premature cervical dilation, chorioamniotic membrane rupture and preterm labour. While up to 40% of preterm labour cases arise from microbial infection, the mechanisms underlying this process are not understood. Notably, select members of the vaginal microbiome have been significantly associated with preterm labour including Sneathia, Prevotella and Porphyromonas species.

The Effect of Topical and Oral Phenytoin on Increasing Wound Healing of Enterocutaneus Fistula in Wistar Rat

BACKGROUND: Enterocutaneous fistula is an abnormal connection between gastrointestinal tract with skin or wound and studies showed low numbers of spontaneous fistula closure (20–30%) despite proper wound treatment and nutrition with high morbidity and mortality rate in fistula repair operation. Phenytoin, commonly used as an anticonvulsant, has shown its effects on wound healing process such as promoting fibroblast activities, formation of granulation tissue, reduction of collagenase activities, increasing collagen production, and other connective tissue components, reducing bacterial colonization, and reducing wound exudate. AIM: To assess the effects of phenytoin in enterocutaneous fistula wound healing in Wistar rats (Ratus norvegicus). MATERIALS AND METHODS: This is an experimental study with randomized post-test only design on 20 Winstar rats. A 5 mm enterocutaneous fistula was made on the rat’s stomach and the rats were assigned randomly into three groups: K (control), P1 (10% phenytoin ointment), and P2 (0.03 mg/g oral phenytoin). The groups were terminated on day 7 and wound histological slides were made. The data were analyzed using SPSS software. RESULTS: The delta diameter is highest in P1 group (mean + SD 0.928 + 0.078), followed by P2 (mean + SD 0.770 + 0.145), and control (mean + SD 0.411 + 0.120). There is a significant difference, p &lt; 0.05 (0.000), between P1 and P2, indicating that oral phenytoin is more effective in collagen formation than topical phenytoin. There is no significant difference between P1 and P2, p &lt; 0.05 (0.269), indicating that oral phenytoin is not more effective than topical phenytoin in granulation tissue in enterocutaneous fistula in Wistar rats. CONCLUSION: Administration of topical and oral phenytoin was effective in increasing granulation tissue thickness, increasing collagen amount in wound tissue, and reducing the diameter of enterocutaneous fistulas in Wistar rats compared with control on the seventh day.

Метаболизм соединительной ткани у больных с гнойными осложнениями после остеосинтеза отломков длинных костей

Остеосинтез (osteosynthesis; греч. osteon (кость) + synthesis (соединение)) — это хирургическое соединение отломков костей в правильном положении с целью стабильной фиксации до полной их консолидации (костного сращения) и восстановления целости и функции кости. Обследовано 34 больных в возрасте 17–58 лет с гнойными осложнениями после остеосинтеза при переломах длинных костей, в том числе у 29 мужчин и 5 женщин. В сыворотке кpови больных определяли следующие биохимические показатели: активность коллагеназы, фpакции гидpоксипpолина, гликозаминогликаны, кальций, фосфор и электролиты (Na+, K+, Ca++).Результаты исследования показывают, что у больных с гнойными осложнениями после остеосинтеза длинных костей при рецидиве воспалительного процесса усиливается катаболическая фаза метаболизма основных компонентов органической основы костной ткани. Об этом свидетельствуют показатели, отражающие синтетическую и катаболическую фазы метаболизма основных компонентов соединительной ткани — коллагена и гликозаминогликанов. Данные, полученные при исследовании минерального обмена, отражают нарушения минерализации костной ткани, что подтверждается показателями содержания общего кальция, ионизированного кальция, фосфора и электролитов.

Phytochemical Analysis and Antioxidant, Antimicrobial, and Antiaging Activities of Ethanolic Seed Extracts of Four Mucuna Species

The investigation into promising botanical materials for natural cosmetics is expanding due to environmental and health awareness. Here, we aimed to evaluate the phytochemical substances and the potential skin-related pharmacological activities of four Mucuna seeds, namely M. gigantea (Willd.) DC. (MGG), M. interrupta Gagnep. (MIT), M. monosperma Wight (MMM), and M. pruriens (L.) DC. (MPR), belonging to the Fabaceae family. In methodology, the Mucuna seeds were authenticated using morphological and molecular approaches. L-DOPA, phenolics, and flavonoid content, incorporated with HPLC and GC–MS fingerprinting analyses, were determined. Then, skin-related antimicrobial, antioxidant, and antiaging activities were determined. The results revealed that MPR showed the highest L-DOPA content (75.94 mg/100 mg extract), whereas MGG exhibited the highest phenolic and flavonoid content (56.73 ± 0.62 mg gallic/g extract and 1030.11 ± 3.97 mg quercetin/g extract, respectively). Only MMM and MPR could inhibit all of S. aureus, S. epidermidis, and C. albicans, but no sample could inhibit C. acnes. Furthermore, all samples demonstrated antioxidant activity. Interestingly, all Mucuna samples exhibited strong collagenase, elastase, and hyaluronidase inhibitory activities. We conclude that the ethanolic extracts of four Mucuna seeds are probably advantageous in the development of skincare cosmeceutical products.

Mitigating the negative impacts of marine invasive species – Sargassum muticum - a key seaweed for skincare products development

Sargassum muticum is a highly invasive species, threatening marine biodiversity worldwide. One strategy to reduce marine invaders' impacts is to promote their use as valuable biomass for new products development. On the other hand, there is a rising conscience of natural compounds importance as health promoters. The present work was designed to sustainably use the marine invasive seaweed S. muticum collected off the Portuguese shore for novel skincare products. The antioxidant, anti-enzymatic (collagenase, elastase, hyaluronidase, tyrosinase), antimicrobial (Staphylococcus epidermidis, Cutibacterium acnes, Malassezia furfur), photoprotective, and anti-inflammatory properties of five fractions (F1–F5) obtained by a sequential extraction of S. muticum were evaluated. The diethyl ether fraction (F2) demonstrated the most promising results, with the highest antioxidant and photoprotective capacity, reducing reactive oxygen species (ROS) production promoted by UVA and UVB radiation in 3T3 cells. On the other hand, the ethyl acetate fraction (F3) exhibited the highest anti-enzymatic capacity, inhibiting the activities of collagenase, hyaluronidase and tyrosinase (IC50 of 97.5, 23.7 and 72.3 μg/mL, respectively). Moreover, fractions from S. muticum showed anti-inflammatory potential by reducing tumor necrosis factor – α and interleukin-6 release. A chemical screening by 1H NMR of S. muticum fractions evidenced signals that can be attributed to the presence of different chemical classes, including lipids, pigments, amino acids, polyphenols, and sugars, being related to the observed multitarget properties. This work highlights a strategic valorisation of S. muticum as a source of treasured ingredients for skincare applications.

Clinical Anti-aging Efficacy of Propolis Polymeric Nanoparticles Prepared by a Temperature-induced Phase Transition Method.

Collagen forms a dermal matrix in the skin. Biosynthesis and decomposition of collagen are the major processes in skin aging. Propolis is rich in flavonoids and phenolic compounds, which are known to be effective in preventing skin aging, including the enhancement of fibroblast proliferation, activation, and growth capacity. The aim of this study was to develop a poorly soluble propolis extract as an active ingredient in cosmetic products for anti-aging efficacy. Polymeric nanoparticles containing propolis extract, polyethylene glycol 400, and poloxamer 407 were prepared via a temperature-induced phase transition method. The particle size of the polymeric nanoparticles was approximately 20.75nm. The results of an in vitro procollagen type I carboxy-terminal peptide assay and a matrix metalloproteinase-1 inhibition assay showed that the polymeric nanoparticles increased collagen production by 19.81%-24.59% compared to blank (p<0.05), and significantly reduced intracellular collagenase activity by 7.46%-31.52% compared to blank (p<0.05). In a clinical trial, polymeric nanoparticles in a cosmetic formulation were applied around the eyes of 24 female subjects for 8weeks. Five skin parameters were significantly improved after the application of the test ampoule. Visual evaluation using the Global Photo Damage Score showed a significant reduction in wrinkles after the application of the test ampoules (p<0.001). This study outlines the development of stable polymeric nanoparticles containing poorly soluble propolis in a cosmetic formulation, and its efficacy in wrinkle improvement. The developed polymeric nanoparticles were effective for alleviating wrinkles and can be used for pharmaceutical applications that utilize propolis as antiseptic, anti-inflammatory, antimycotic, antifungal, antibacterial, antiulcer, anticancer, and immunomodulatory agents.

Characterization and application of a novel halotolerant protease with no collagenase activity for cleaner dehairing of goatskin

The current study focused on the substitution of toxic Na2S with bio-enzyme G3726, a novel serine protease heterologously expressed by Bacillus subtilis strain SCK6. This protease was purified to electrophoretic homogeneity with 39.43 % recovery, 7.03-fold purification and an estimated molecular mass of 36 kDa. The maximal activity was achieved at pH 8, 75 °C and 40 g L−1 NaCl, and more than 70 % of the activity was retained across the broad salinity range of 0–200 g L−1. G3726 was activated by Sr2+ and Ca2+, displayed considerable stability to benzene and ethanediol, and exhibited excellent compatibility with Tweens (20 and 80) and Triton X-100. Simultaneously, the 3D structure of G3726 was constructed by homology modeling, and interacted well with the cementing substances glycoprotein and proteoglycan in the hair pores. Moreover, G3726 showed no collagenase activity, and could thoroughly dehair goatskin within 4 h without any damage, which was better than industrial enzyme AS1.398 and chemical Na2S. As compared to Na2S, G3726 enzymatic dehairing showed reduction in BOD5, COD and TSS by 53.81, 52.78 and 65.52 %, respectively, endowing the leather better physical properties. These outstanding findings indicate that protease G3726 may have application in dehairing for environment-friendly leather processing.

83 - Verification of Collagenase Production by Streptococcus Mutans

The oral environment contains multiple collagenases which are believed to play a role in dentin caries formation, as collagen is one of the major components of dentin. In the laboratory, biofilm and collagenase-containing chemical models have been used to study dental degradation. However, the numerous collagenase sources in the oral cavity pose a challenge when it comes to comparing in vitro and in vivo data. Streptococcus mutans, one of the important cariogenic oral streptococcu, contains genes for at least two collagenases (SMU_759 and SMU_761), which have been shown to be expressed in samples collected from active caries. Here, we investigate if the collagenases produced by S. mutans contribute to dentin degradation. S. mutans UA159 (ATCC700610) was cultured in BHI with/without KHCO3 using different concentrations of sucrose (0%, 0.05%, 0.2%, and 1%) at 37 C and 5% CO2 for 48 hrs. The pH of the media at 48 hrs was measured by using pH test paper strips. RNA from S. mutans biofilms cultured without KHCO3 was collected to measure collagenase gene expression by RT qPCR. To measure collagenase activity, KHCO3-buffered BHI was used for culturing to avoid collagenase denaturing caused by low pH. Supernatants of the same culture were centrifuged, filter sterilized and dialyzed, and exposed to a commercially available colorimetric assay kit. Absorbance readings were collected every 20 min for 16 hrs. With KHCO3 buffering, the pH of the media containing 48-hrs S. mutans cultured with 0%, 0.05%, 0.2%, and 1% sucrose were around 7.6, 6.5, 6.0, and 5.5, respectively. The corresponding pH values of media without KHCO3 were around 7.0, 5.5, 5.0, and 4.5. Addition of 1% W/V sucrose in BHI led to an approximately 20-fold increase in the expression of both SMU_759 and 761 collagenases compared to BHI only. Collagenase expression increased by less than 3-fold when 0.05% and 0.2% W/V sucrose was added. Although quite slow, a continuous reduction in absorbance was observed in the 16-hr collagenase activity test, indicating collagenase activity of the media containing S. mutans. The collagenase activities of the samples with 0%, 0.05%, 0.2%, and 1% sucrose were: 0.00106, 0.000679, 0.000679, and 0.000931 U/ml, respectively. Gene expression and collagenase activity test showed that S. mutans produced active collagenase(s) in response to sucrose during the 48 hrs of culturing, which supports the idea that S. mutans contributes to dentin degradation.

Efficiency of topical phenytoin in diabetic foot ulcer care: A randomized control trial

Aim: To study the efficiency of topical phenytoin on healing in diabetic foot ulcer grades 1 and 2. Methods: Totally, 60 patients in each group with diabetic foot ulcers of more than 4 weeks duration without any co-morbid conditions and ulcers belonging to Grade 1 and 2 as per Meggitt – Wagner’s diabetic foot ulcers were inducted into the study. The whole sample population was divided into two equal and comparable groups based on willingness for undergoing topical phenytoin therapy for the wound. Those who were not willing were subjected to conventional wound care, forming the control group. Antibiotics are given to both groups as per culture reports. Results: In the study group, 13 patients had granulation tissue at 14 days of less than 75%, and 17 patients had granulation tissue at 14 days of greater than 75%. In the control group mean value of the size of the ulcer initially is 8.76cm and in 7 days is 7.12 cm and in 14 days is 5.86 cm. In the study group mean value of the size of the ulcer initially is 9.03 cm and in 7 days is 5.63 cm and in 14days is 3.3 cm. Conclusion: Phenytoin enhances wound healing via the following mechanisms: stimulation of fibroblast proliferation, enhancement of granulation tissue development, and inhibition of collagenase activity.

One-pot synthesis and biochemical characterization of a magnetic collagenase nanoflower and evaluation of its biotechnological applications

Recently, hierarchical magnetic enzyme nanoflowers have been found extensive attention for efficient enzyme immobilization due to high surface area, low mass transfer limitations, active site accessibility, promotion of the enzymatic performance, and facile reusing. Herein, we report the purification of the Bacillus collagenase and then synthesis of magnetic cross-linked collagenase-metal hybrid nanoflowers (mcCNFs). The catalytic efficiency (kcat/Km) value of the immobilized collagenase was 2.2 times more than that of the free collagenase. The collagenase activity of mcCNFs enhanced about 2.9 and 4.6 at 85 and 90 °C, respectively, compared to free collagenase. Thermal stability of mcCNFs increased about 31% and 24% after 3 h of incubation at 50 and 60 °C, respectively. After 10 cycles of reusing, the mCNFs collagenase showed 83% of its initial activity. Results showed that the mcCNFs revealed 1.4 times more activity than the free collagenase in 0.16% protein waste. Furthermore, the hydrolysis value of chicken pie protein wastes by the immobilized enzyme obtained 4 times more than the free collagenase after 240 min incubation at 40 °C. Finally, our results showed that the construction of mcCNFs is an efficient method to increase the enzymatic performance and has excessive potential for the hydrolysis of protein wastes in the food industry.