A Novel Gelatinase from Marine Flocculibacter collagenilyticus SM1988: Characterization and Potential Application in Collagen Oligopeptide-Rich Hydrolysate Preparation

Although the S8 family in the MEROPS database contains many peptidases, only a few S8 peptidases have been applied in the preparation of bioactive oligopeptides. Bovine bone collagen is a good source for preparing collagen oligopeptides, but has been so far rarely applied in collagen peptide preparation. Here, we characterized a novel S8 gelatinase, Aa2_1884, from marine bacterium Flocculibacter collagenilyticus SM1988T, and evaluated its potential application in the preparation of collagen oligopeptides from bovine bone collagen. Aa2_1884 is a multimodular S8 peptidase with a distinct domain architecture from other reported peptidases. The recombinant Aa2_1884 over-expressed in Escherichia coli showed high activity toward gelatin and denatured collagens, but no activity toward natural collagens, indicating that Aa2_1884 is a gelatinase. To evaluate the potential of Aa2_1884 in the preparation of collagen oligopeptides from bovine bone collagen, three enzymatic hydrolysis parameters, hydrolysis temperature, hydrolysis time and enzyme-substrate ratio (E/S), were optimized by single factor experiments, and the optimal hydrolysis conditions were determined to be reaction at 60 ℃ for 3 h with an E/S of 400 U/g. Under these conditions, the hydrolysis efficiency of bovine bone collagen by Aa2_1884 reached 95.3%. The resultant hydrolysate contained 97.8% peptides, in which peptides with a molecular weight lower than 1000 Da and 500 Da accounted for 55.1% and 39.5%, respectively, indicating that the hydrolysate was rich in oligopeptides. These results indicate that Aa2_1884 likely has a promising potential application in the preparation of collagen oligopeptide-rich hydrolysate from bovine bone collagen, which may provide a feasible way for the high-value utilization of bovine bone collagen.

Forced Degradation Study for Simultaneous Quantification of Aspirin and Omeprazole in Pharmaceutical Dosage form by RP-HPLC

Aims: To study force degradation of aspirin and omeprazole simultaneously by RP-HPLC method Study design: RP-HPLC method was used to measure % degradation. Place and Duration of Study: Study was carried out at center of excellence, G.I.D.C., vapi-396195, Gujarat, India between June 2019 to march 2020. Methodology: A force degradation study of aspirin and omeprazole was carried out simultaneously. The drugs were subjected to various degradation conditions like hydrolysis by acid and base, Oxidative degradation, and thermal degradation study. Results: For acidic condition, the degradation was found to be 32.63 % for aspirin and 61.64 % for omeprazole. For basic condition, the degradation was found to be 10.17 % for aspirin and 4.29 % for omeprazole. By oxidative hydrolysis, the aspirin was degraded by 15.48 % and omeprazole was degraded by 26.38 %. By thermal degradation, 0.37 % degradation was observed for aspirin and 4.32 % degradation for omeprazole. Conclusion: In this proposed method the retention time for drug is less than 8 min, which is less then available method. For omeprazole, strong degradation was observed in acidic conditions and mild in basic hydrolysis conditions. For aspirin, more degradation was observed in basic conditions than acidic hydrolysis. Both drugs were degraded in oxidative conditions using 3% H2O2. Omeprazole degraded more than aspirin by dry heat degradation. The method was successfully applied for the quantitative determination of both Active Pharmaceutical Ingredients.

Protease Catalyzed Production of Spent Hen Meat Hydrolysate Powder for Health Food Applications

Whole spent hen meat of Indian commercial layer bird (BV-300 breed) was enzymatically hydrolyzed using Flavourzyme® derived from Aspergillus oryzae. Different time, temperature, and pH combinations generated through response surface methodology (RSM) were tested to find the optimal hydrolysis condition at which maximum antioxidant potential and degree of hydrolysis can be achieved. Hydrolysis for 30 min at a temperature of 53.9°C and pH of 6.56 was found suitable for achieving high degree of hydrolysis and antioxidant activity. Antioxidant potential at optimized conditions was estimated at 93.26% by DPPH radical scavenging assay and 2.32 mM TEAC by FRAP assay. Amino acid profiling of the hydrolysate correlated very well with SDS-PAGE profiling. SDS-PAGE results confirmed that 30 min hydrolysis time was enough to produce low molecular weight peptides (2–5 kDa) with high antioxidant potential. Antioxidant rich Indian spent hen meat hydrolysate powder was economically produced using spray drying. Sensory analysis revealed that 10% hydrolysate powder had satisfactory overall acceptability and has potential to be used in health/functional foods at this concentration. This is the first study wherein optimum hydrolysis conditions for Indian spent hen meat have been reported.

Industrial Gelatin from Leather Chrome Shavings Wastes

A step towards minimizing the environmental pollution of leather tanning , leather chrome shavings wastes were treated with Li2CO3 to extract technical or industrial gelatin as an added value material. Isolation and characterization of gelatin obtained from chrome-tanned shavings were done. The alkali hydrolysis products obtained, showed good physical and chemical properties in terms of gel strength, swelling and thermal stability. The optimum hydrolysis conditions using Li2CO3 were found to be 5 hr. extraction at 80°C, swelling time of one day and pH 9.5. The yield was over one third of the original starting waste material.

Preparation of High-purity Alumina via Hydrolysis of Aluminum Isopropoxide after Desilication with Lanthanum Oxide

High-purity alumina refers to ultra-fine alumina powder with a purity exceeding 99.99% and a uniform particle size. This material exhibits excellent corrosion resistance, high-temperature resistance, wear resistance, and oxidation resistance. Owing to the high silicon content of alumina prepared by means of the alcohol-aluminum hydrolysis method, the purity of the alumina is often unsatisfactory. Therefore, in this work, a new method for adding lanthanum oxide to isopropanol in the early aluminum isopropoxide synthesis stage is proposed. When lanthanum oxide was added, the silicon content of the precursor aluminum isopropoxide decreased to 0.0051%.Remove calcium, sodium, magnesium and other impurities by cleaning with hydrochloric acid under an ultrasonic field. The optimal hydrolysis conditions were determined as follows: hydrolysis temperature: 55, hydrolysate concentration: 80%, water to alkoxide ratio: 6:1. The alumina precursor calcined at 1200 yielded a high-purity alumina with a purity level of more than 99.99%, and the particle size reaches 2.037 μm.

Synthesis of Cellulose Nanocrystals (CNCs) from Brewer’s Spent Grain Using Acid Hydrolysis: Characterization and Optimization

This study is aimed at utilizing brewery’s spent grain (BSG) byproduct for the synthesis of cellulose nanocrystals (CNCs) using acid hydrolysis and optimizing the hydrolysis parameters (hydrolysis time, temperature, liquid-solid ratio, and acid concentration). Alkali and bleaching treatment were done to remove hemicellulose and lignin from BSG. Optimization process was performed using central composite design (CCD) to obtain optimum value of cellulose nanocrystal (CNC) yield. The maximum cellulose nanocrystal (CNC) yield of 43.24% was obtained at optimum hydrolysis conditions of 50°C, 51 wt% acid concentration, 41 min, and liquid-solid ratio of 19 ml/g. The raw brewery spent grain; alkali-treated fiber, bleached fiber, and obtained CNC were characterized using scanning electron microscopy (SEM), XRD, particle analyzer, FTIR, and differential scanning calorimeter (DSC). The characterization results indicated that the obtained cellulose nanocrystal (CNC) has rod-like whisker shape with crystallinity of 76.3% and an average particle size of 309.4 nm.

Bioethanol Production from Sugarcane Press-Mud: Assessment of the Fermentation Conditions to Reduce Fusel Alcohol

Within a biorefinery context, bioethanol is a promising platform molecule since it can be used as raw material to produce a wide spectrum of valuable industrial products such as H2 and light olefins. However, the presence of impurities limits the conversion of bioethanol in these products. Herein, we aimed to determine the proper pretreatment and fermentation conditions to yield bioethanol with a low content of impurities, such as 3-methyl-1-butanol, by using sugarcane press-mud as feedstock. To do so, a BoxBehnken methodology was employed to select proper pretreatment and fermentation conditions. Factors assessed were temperature, stirring, and pH during fermentation of hydrolysates coming from two different pretreatment methods named as hydrothermal and acid hydrolysis. Results showed that the fermentation temperature should be kept between 26–30 °C to assure at least 91 g/L ethanol. The fusel alcohol content would be reduced by 22% at 30 °C, pH = 4.5, and 200 rpm if sugarcane press-mud is pretreated under acid hydrolysis conditions (T = 130 °C, t = 1 h, 16 g HNO3/kg solid). Further studies should aim to integrate these conditions within a biorefinery concept to yield valuable products such as H2 and ethylene.