Invitro bioprocessing of corn as poultry feed additive by the influence of carbohydrate hydrolyzing metagenome derived enzyme cocktail

The carbohydrate-hydrolyzing enzymes play a crucial role in increasing the phenolic content and nutritional properties of polysaccharides substrate, essential for cost-effective industrial applications. Also, improving the feed efficiency of poultry is essential to achieve significant economic benefits. The current study introduced a novel thermostable metagenome-derived xylanase named PersiXyn8 and investigated its synergistic effect with previously reported α-amylase (PersiAmy3) to enhance poultry feed utilization. The potential of the enzyme cocktail in the degradation of poultry feed was analyzed and showed 346.73 mg/g poultry feed reducing sugar after 72 h of hydrolysis. Next, the impact of solid-state fermentation on corn quality was investigated in the presence and absence of enzymes. The phenolic content increased from 36.60 mg/g GAE in control sample to 68.23 mg/g in the presence of enzymes. In addition, the enzyme-treated sample showed the highest reducing power OD 700 of 0.217 and the most potent radical scavenging activity against ABTS (40.36%) and DPPH (45.21%) radicals. Moreover, the protein and ash contents of the fermented corn increased by 4.88% and 6.46%, respectively. These results confirmed the potential of the carbohydrate-hydrolyzing enzymes cocktail as a low-cost treatment for improving the phenolic content, antioxidant activity, and nutritional values of corn for supplementation of corn-based poultry feed.

Development of Lignocellulase Enzyme Cocktail—A Logical Use of Statistical Experimental Design Techniques

The derivation of reduced sugars from lignocellulosic biomass requires an optimum blend of cellulolytic enzymes and reaction conditions that favour high sugar yield. In this respect, statistical design of experiment strategies become useful, but the technique is often misguided such that enzyme redundancy becomes overlooked. Herein, we demonstrate a systematic approach that involves simplex lattice mixture design and central composite design for optimizing enzyme cocktails for the saccharification of lignocellulosic biomass. The simplex lattice mixture design yielded 0.3333: 0.3333: 0.3333 of Celluclast (5%), Hemicellulase (5%) and Laccase (2%), respectively, as the optimum enzyme blend (volume) ratio for the saccharification of hydrothermally pretreated empty palm fruit bunch (EPFB, 10% solid loading). A subsequent application of central composite design resulted in 40 oC, pH 6 and 24 hrs as the optimum saccharification conditions. The individual Celluclast (5%) and Hemicellulase (5%) yielded a reduced sugar equivalence (RSE) of 1.77 mg/mL and 1.67 mg/mL, respectively. However, the blended enzyme cocktail upon subjection to simplex lattice mixture design and subsequent central composite design yielded an RSE of 2.215 mg/mL and 2.431 mg/mL, respectively. The overall results exemplify the significance of enzyme synergism in lignocellulosic biomass saccharification. The approach herein is intended as an easy-to-copy plan for optimizing enzyme cocktails.

Effect of Soluble Phenolic Compounds From Hydrothermally Pretreated Wheat Straw on Cellulose Degrading Enzymes

Background: Hydrothermal methods are commonly applied in pretreatment of lignocellulose for enhanced enzymatic hydrolysis and further conversion to biofuels and chemicals. The pretreatment partially disassemble and solubilize cell wall polymers. Besides mono- and oligosaccharides, the soluble degradation products include various phenolic compounds, which may affect the efficiency of enzymatic hydrolysis. Results: The phenolic compounds were isolated from pretreatment liquor and their effects on cellulolytic enzymes were investigated. The major enzymes in crystalline cellulose degradation, cellobiohydrolases, were inhibited by the oligophenolics and phenolic-carbohydrate conjugates the latter of which could be mitigated by other enzymes, i.e. xylanases and endoglucanases. Hydrolytic activity of a commercial enzyme cocktail containing lytic polysaccharide monooxygenase (LPMO) was enhanced in presence of low concentrations of phenolics.Conclusions: The effects of the pretreatment liquor phenolics is dependent on enzyme concentration and type of the phenolic compounds. For optimized performance, the pretreatment conditions and enzyme cocktail composition should be designed to promote oxidative activities for enhanced hydrolysis, degradation of the oligosaccharides linked to phenolics and resistance to oligophenolic compounds.

Proteases Production and Chitin Preparation from the Liquid Fermentation of Chitinous Fishery By-Products by Paenibacillus elgii

Chitinous fishery by-products have great application in the production of various bioactive compounds. In this study, Paenibacillus elgii TKU051, a protease-producing bacterial strain, was isolated using a medium containing 1% squid pens powder (SPP) as the sole carbon/nitrogen (C/N) source. P. elgii TKU051 was found to produce at least four proteases with molecular weights of 100 kDa, 57 kDa, 43 kDa, and 34 kDa (determined by the gelatin zymography method). A P. elgii TkU051 crude enzyme cocktail was optimally active at pH 6–7 and 60 °C. The 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and α-glucosidase inhibitory activity of the hydrolysates obtained from the hydrolysis of shrimp shell powder, shrimp head powder, shrimp meat powder, fish head powder and soya bean powder catalyzed by the P. elgii TkU051 crude enzyme cocktail were also evaluated. P. elgii TKU051 exhibited a high deproteinization capacity (over 94%) on different kinds of shrimp waste (shrimp heads and shells; fresh and cooked shrimp waste; shrimp waste dried by oven and lyophilizer), and the Fourier-transform infrared spectroscopy profile of the chitin obtained from the deproteinization process displayed the characteristic of chitin. Finally, the obtained chitin exhibited an effect comparable to commercial chitin in terms of adsorption against Congo Red (90.48% and 90.91%, respectively). Thus, P. elgii TKU051 showed potential in the reclamation of chitinous fishery by-products for proteases production and chitin extraction.