Production of Enzymatic Extract with High Cellulolytic and Oxidative Activities by Co-Culture of Trichoderma reesei and Panus lecomtei

Abstract

This work aimed to produce enzymatic fungi extracts with hydrolytic and oxidative activities to hydrolyze lignocellulosic biomasses efficiently. For this, the fungi Trichoderma reesei and Panus lecomtei were co-cultured using the vegetable biomasses oil palm decanter cake, wheat bran, and cottonseed cake as substrates in submerged fermentation. T. reesei and P. lecomtei showed partially compatible positive interaction on plates. The co-cultures respond positively to variations of temperature and inoculum interval, generating extracts responsible for higher hydrolysis yield when grown at 25 °C, and P. lecomtei is inoculated 24 h after T. reesei. The enzymatic extract production of co-cultures was also improved by modifying the components of the initial media and evaluating enzymatic activities, hydrolysis of sugarcane bagasse pretreated by autohydrolysis and ethanol production as a response. Five culture media were evaluated with variations in the composition of nutritional elements, minerals and substrates. The best extract showed a maximum cellulose hydrolysis efficiency of 68.7% compared with 44.8% of the initial medium. The ethanolic fermentation of hydrolysates obtained by co-culture extracts showed higher ethanol yields than monocultures. This work demonstrates the use of fungi co-cultures to produce enzymatic extracts composed of cellulolytic, hemicellulolytic, and ligninolytic enzymes complexes, which allow hydrolyzing pretreated lignocellulosic biomass with high efficiency, generating hydrolysates that are easier fermented by yeast.

Crude Enzyme Concentrate of Filamentous Fungus Hydrolyzed Chitosan to Obtain Oligomers of Different Sizes

“…Initially, two culture media were evaluated for better enzymatic activity, established as endoglucanase activity [26]. Two media were used, modified Czapeck (C zm ): carboxymethylcellulose (CMC) 10.0 g/L, sodium nitrate 3.0 g/L, potassium phosphate 1.0 g/L, potassium chloride 0.5 g/L, magnesium sulfate 0.5 g/L, and ferrous sulfate 0.01 g/L; and CPY: carboxymethylcellulose (CMC) 10.0 g/L, peptone 1.0 g/L, and yeast extract 20.0 g/L.…”

Improvement of Laccase Activity in Co-Culture of Panus lecomtei and Sporidiobolus pararoseus and Its Application as an Enzymatic Additive in Biomass Hydrolysis and Dye Decolorization

“…For the cultivation, a liquid media based on the work of Peláez and collaborators [5] containing urea (0.45 g/L), yeast extract (0.75 g/L), bacteriological peptone (0.13 g/L), (NH 4 ) 2 SO 4 (2.1 g/L), KH 2 PO 4 (2.0 g/L), CaCl 2 (0.2 g/L), MgSO 4 .7H 2 O (0.45 g/L), CuSO 4 .5H 2 O (0.43 g/L), ZnSO 4 .7H 2 O (1.4 mg/L), MnSO 4 .H 2 O (1.04 mg/L) was prepared. The culture media was supplemented with the biomass OPDC (25 g/L), CS (50 g/L), and WB (20 g/L) as carbon sources.…”

“…After that, it was resuspended in water until an optical density (OD) close to 1.0 (1 × 10 6 UFC/mL) was measured at 600 nm. In co-culture, 5 mycelial discs of fully colonized agar of P. lecomtei were inoculated in the media based on the work of Peláez et al [5] and as described in Section 2.2.1. After 120 h, each flask was inoculated with 1 mL (2% v/v) of S. pararoseus inoculum.…”

“…The calculations of hydrolysis efficiency (%) were based on the conversion rate of cellulose and hemicellulose into glucose and xylose [18]. All the equations can be found in detail in the work of Peláez et al (2022) [5]. Compositional analysis of PSB was determined following the protocol recommended by the National Renewable Energy Laboratory [19].…”

“…Enzyme production is one of the main applications of interspecific interactions of fungi in synthetic culture media. In competitive relationships, when two or more fungi interact in direct contact, numerous changes are noticed at the morphological level or in the secretion of metabolites and enzymes [4,5]. Several studies have shown that co-cultures of two fungi with different enzymatic profiles, such as Trichoderma spp and white-rot fungi, can act in synergy to produce improved versions of these extracts, rich in cellulases and laccases [6].…”