Bioprocessing of Two Crop Residues for Animal Feeding into a High-Yield Lovastatin Feed Supplement.

This work aimed to evaluate the lovastatin (Lv) production by solid-state fermentation (SSF) from selected crop residues, considering the post-fermented residues as feed supplements for ruminants. The SSF was performed with two substrates (wheat bran and oat straw) and two A. terreus strains (CDBB H-194 and CDBB H-1976). The Lv yield, proximate analysis, and organic compounds by GC-MS in the post-fermented residues were assessed. The combination of the CDBB H-194 strain with oat straw at 16 d of incubation time showed the highest Lv yield (23.8 mg/g DM fed) and the corresponding degradation efficiency of hemicellulose + cellulose was low to moderate (24.1%). The other three treatments showed final Lv concentrations in decreasing order of 9.1, 6.8, and 5.67 mg/g DM fed for the oat straw + CDBB H-1976, wheat bran + CDBB H-194, and wheat bran + CDBB H-1976, respectively. An analysis of variance of the 22 factorial experiment of Lv showed a strong significant interaction between the strain and substrate factors. The kinetic of Lv production adequately fitted a zero-order model in the four treatments. GC-MS analysis identified only a couple of compounds from the residues fermented by A. terreus CDBB H-194 (1,3-dipalmitin trimethylsilyl ether in the fermented oat straw and stearic acid hydrazide in the fermented wheat bran) that could negatively affect ruminal bacteria and fungi. Solid-state fermentation of oat straw with CDBB H-194 deserves further investigation due to its high yield of Lv; low dietary proportions of this post-fermented oat straw could be used as an Lv-carrier supplement for rumen methane mitigation.

Fermentation performance of a Mexican native Clavispora lusitaniae strain for xylitol and ethanol production from xylose, glucose and cellobiose.

Lignocellulose hydrolysates are rich in fermentable sugars such as xylose, cellobiose and glucose, with high potential in the biotechnology industry to obtain bioproducts of higher economic value. Thus, it is important to search for and study new yeast strains that co-consume these sugars to achieve better yields and productivity in the processes. The yeast Clavispora lusitaniae CDBB-L-2031, a native strain isolated from mezcal must, was studied under various culture conditions to potentially produce ethanol and xylitol due to its ability to assimilate xylose, cellobiose and glucose. This yeast produced ethanol under microaerobic conditions with yields of 0.451 gethanol/gglucose and 0.344 gethanol/gcellobiose, when grown on 1% glucose or cellobiose, respectively. In mixtures (0.5% each) of glucose:xylose and glucose:xylose:cellobiose the yields were 0.367 gethanol/gGX and 0. 380 gethanol/gGXC, respectively. Likewise, in identical conditions, C. lusitaniae produced xylitol from xylose with a yield of 0.421 gxylitol/gxylose. In 5% glucose or xylose, this yeast had better ethanol and xylitol titers and yields, respectively. However, glucose negatively affected xylitol production in the mixture of both sugars (3% each), producing only ethanol. Xylose reductase (XR) and xylitol dehydrogenase (XDH) activities were evaluated in cultures growing on xylose or glucose, obtaining the highest values in cultures on xylose at 8 h (25.9 and 6.22 mU/mg, respectively). While in glucose cultures, XR and XDH activities were detected once this substrate was consumed (4.06 and 3.32 mU/mg, respectively). Finally, the XYL1 and XYL2 genes encoding xylose reductase and xylitol dehydrogenase, respectively, were up-regulated by xylose, whereas glucose down-regulated their expression.

Lovastatin as a supplement to mitigate rumen methanogenesis: an overview.

Methane from enteric fermentation is the gas with the greatest environmental impact emitted by ruminants. Lovastatin (Lv) addition to feedstocks could be a strategy to mitigate rumen methane emissions via decreasing the population of methanogenic archaea (MA). Thus, this paper provides the first overview of the effects of Lv supplementation, focusing on the inhibition of methane production, rumen microbiota, and ruminal fermentation. Results indicated that Lv treatment had a strong anti-methanogenic effect on pure strains of MA. However, there are uncertainties from in vitro rumen fermentation trials with complex substrates and rumen inoculum.Solid-state fermentation (SSF) has emerged as a cost-effective option to produce Lv. In this way, SSF of agricultural residues as an Lv-carrier supplement in sheep and goats demonstrated a consistent decrease in ruminal methane emissions. The experimental evidence for in vitro conditions showed that Lv did not affect the volatile fatty acids (VFA). However, in vivo experiments demonstrated that the production of VFA was decreased. Lv did not negatively affect the digestibility of dry matter during in vitro and in vivo methods, and there is even evidence that it can induce an increase in digestibility. Regarding the rumen microbiota, populations of MA were reduced, and no differences were detected in alpha and beta diversity associated with Lv treatment. However, some changes in the relative abundance of the microbiota were induced. Further studies are recommended on: (i) Lv biodegradation products and stability, as well as its adsorption onto the solid matter in the rumen, to gain more insight on the "available" or effective Lv concentration; and (ii) to determine whether the effect of Lv on ruminal fermentation also depends on the feed composition and different ruminants.

Biological synthesis of iron nanoparticles using hydrolysates from a waste-based biorefinery.

The purpose of this work was to produce iron nanoparticles (Fe-NP) by microbial pathway from anaerobic bacteria grown in anaerobic fluidized bed reactors (AnFBRs) that constitute a new stage of a waste-based biorefinery. Bioparticles from biological fluidized bed reactors from a biorefinery of organic fraction of municipal solid wastes (that produces hydrolysates rich in reducing sugars) were nanodecorated (embedded nanobioparticle or nanodecorated bioparticle, ENBP) by biological reduction of iron salts. Factors "origin of bioparticles" (either from hydrogenogenic or methanogenic fluidized bed reactor) and "type of iron precursor salt" (iron chloride or iron citrate) were explored. SEM and high-resolution transmission electron microscopy (HRTEM) showed amorphous distribution of nanoparticles (NP) on the bioparticles surface, although small structures that are nanoparticle-like could be seen in the SEM micrographs. Some agglomeration of NPs was confirmed by DLS. Average NP size was lower in general for NP in ENBP-M than ENBP-H according to HRTEM. The factors did not have a significant influence on the specific surface area of NPs, which was high and in the range 490 to 650 m2 g-1. Analysis by EDS displayed consistent iron concentration 60-65% iron in nanoparticles present in ENBP-M (bioparticles previously grown in methanogenic bioreactor), whereas the iron concentration in NPs present in ENBP-H (bioparticles previously grown in hydrogenogenic bioreactor) was more variable in a range from 8.5 to 62%, depending on the iron salt. X-ray diffraction patterns showed the typical peaks for magnetite at 35° (3 1 1), 43° (4 0 0), and 62° (4 0 0); moreover, siderite diffraction pattern was found at 26° (0 1 2), 38° (1 1 0), and 42° (1 1 3). Results of infrared analysis of ENBP in our work were congruent with presence of magnetite and occasionally siderite determined by XRD analysis as well as presence of both Fe+2 and F+3 (and selected satellite signal peaks) observed by XPS. Our results on the ENBPs hold promise for water treatment, since iron NPs are commonly used in wastewater technologies that treat a wide variety of pollutants. Finally, the biological production of ENBP coupled to a biorefinery could become an environmentally friendly platform for nanomaterial biosynthesis as well as an additional source of revenues for a waste-based biorefinery.

Parametric optimization of domestic wastewater treatment in an activated sludge sequencing batch reactor using response surface methodology.

In this work, the parametric optimization of real domestic wastewater treated in an activated sludge sequencing batch reactor (SBR) was performed by means of the response surface methodology (RSM). The influences of influent organic matter concentration as chemical oxygen demand (CODinf), biomass concentration (Xs) and aeration time (t) on the COD, organic matter removal efficiency as COD (η) and sludge volume index (SVI) were determined to evaluate the performance of activated sludge SBR. The results showed that organic matter efficiency and maximum SVI were obtained at a t of 12 h, 300 mg L-1 of CODinf and 2000 mg L-1 of Xs. The SBR-activated sludge exhibited a η of 73% and an SVI of 119 mL g-1. Both values indicated a very good performance. Furthermore, the COD of the effluent under these conditions complied with Mexican regulations for wastewater discharged into water bodies.

A soft sensor based on online biomass measurements for the glucose estimation and control of fed-batch cultures of Bacillus thuringiensis.

On bioprocess engineering, experimental measurements are always a costly part of the modeling effort; therefore, there is a constant need to develop cheaper, simpler, and more efficient methodologies to exploit the information available. The aim of the present work was to develop a soft sensor with the capacity to perform reliable substrate predictions and control in microbial cultures of the fed-batch type, using mainly microbial growth data. This objective was achieved using dielectric spectroscopy technology for online monitoring of microbial growth and hybrid neural networks for online prediction of substrate concentration. The glucose estimator was integrated to a fuzzy logic controller to control the substrate concentration in a fed-batch experiment. Dielectric spectroscopy is a technology sensitive to the air volume fraction in the culture media and the turbulence generated by the agitation; however, the introduction of a polynomial function for the calibration of the permittivity signal allowed biomass estimations with an approximation error of 2%. The methodology presented in this work was successfully implemented for the glucose prediction and control of a fed-batch culture of Bacillus thuringiensis with an approximation error of 6%.

Expression of a codon-optimized ß-glucosidase from Cellulomonas flavigena PR-22 in Saccharomyces cerevisiae for bioethanol production from cellobiose.

Bioethanol is one of the main biofuels produced from the fermentation of saccharified agricultural waste; however, this technology needs to be optimized for profitability. Because the commonly used ethanologenic yeast strains are unable to assimilate cellobiose, several efforts have been made to express cellulose hydrolytic enzymes in these yeasts to produce ethanol from lignocellulose. The C. flavigenabglA gene encoding ß-glucosidase catalytic subunit was optimized for preferential codon usage in S. cerevisiae. The optimized gene, cloned into the episomal vector pRGP-1, was expressed, which led to the secretion of an active ß-glucosidase in transformants of the S. cerevisiae diploid strain 2-24D. The volumetric and specific extracellular enzymatic activities using pNPG as substrate were 155 IU L-1 and 222 IU g-1, respectively, as detected in the supernatant of the cultures of the S. cerevisiae RP2-BGL transformant strain growing in cellobiose (20 g L-1) as the sole carbon source for 48 h. Ethanol production was 5 g L-1 after 96 h of culture, which represented a yield of 0.41 g g-1 of substrate consumed (12 g L-1), equivalent to 76% of the theoretical yield. The S. cerevisiae RP2-BGL strain expressed the ß-glucosidase extracellularly and produced ethanol from cellobiose, which makes this microorganism suitable for application in ethanol production processes with saccharified lignocellulose.

Bioenergy and bioproducts from municipal organic waste as alternative to landfilling: a comparative life cycle assessment with prospective application to Mexico.

A life cycle assessment (LCA) of a four-stage biorefinery concept, coined H-M-Z-S, that converts 1 t of organic fraction of municipal solid waste (OFMSW) into bioenergy and bioproducts was performed in order to determine whether it could be an alternative to common disposal of OFMSW in landfills in the Mexican reality. The OFMSW is first fermented for hydrogen production, then the fermentates are distributed 40 % to the methane production, 40 % to enzyme production, and 20 % to the saccharification stage. From hydrogen and methane, up to 267 MJ and 204 kWh of gross heat and electricity were produced. The biorefinery proved to be self-sustainable in terms of power (95 kWh net power), but it presented a deficit of energy for heating services (-155 MJ), which was partially alleviated by digesting the wastes from the bioproducts stages (-84 MJ). Compared to landfill, biorefinery showed lower environmental impacts in global warming (down to -128 kg CO2-eq), ozone layer depletion (2.96 × 10-6 kg CFC11-eq), and photochemical oxidation potentials (0.011 kg C2H4-eq). The landfarming of the digestates increased significantly the eutrophication impacts, up to 20 % below the eutrophication from landfilling (1.425 kg PO4-eq). These results suggest that H-M-Z-S biorefinery could be an attractive alternative compared to conventional landfilling for the management of municipal solid wastes, although new alternatives and uses of co-products and wastes should be explored and tested. Moreover, the biorefinery system would benefit from the integration into the market chain of the bioproducts, i.e., enzymes and hydrolysates among others.

Effect of sudden addition of PCE and bioreactor coupling to ZVI filters on performance of fluidized bed bioreactors operated in simultaneous electron acceptor modes.

The present work evaluated the effects of (i) feeding a water contaminated with 80 mg/L PCE to bioreactors seeded with inoculum not acclimated to PCE, (ii) coupling ZVI side filters to bioreactors, and (iii) working in different biological regimes, i.e., simultaneous methanogenic aeration and simultaneous methanogenic-denitrifying regimes, on fluidized bed bioreactor performance. Simultaneous electron acceptors refer to the simultaneous presence of two compounds operating as final electron acceptors in the biological respiratory chain (e.g., use of either O2 or NO3- in combination with a methanogenic environment) in a bioreactor or environmental niche. Four lab-scale, mesophilic, fluidized bed bioreactors (bioreactors) were implemented. Two bioreactors were operated as simultaneous methanogenic-denitrifying (MD) units, whereas the other two were operated in partially aerated methanogenic (PAM) mode. In the first period, all bioreactors received a wastewater with 1 g chemical oxygen demand of methanol per liter (COD-methanol/L). In a second period, all the bioreactors received the wastewater plus 80 mg perchloroethylene (PCE)/L; at the start of period 2, one MD and one PAM were coupled to side sand-zero valent iron filters (ZVI). All bioreactors were inoculated with a microbial consortium not acclimated to PCE. In this work, the performance of the full period 1 and the first 60 days of period 2 is reported and discussed. The COD removal efficiency and the nitrate removal efficiency of the bioreactors essentially did not change between period 1 and period 2, i.e., upon PCE addition. On the contrary, specific methanogenic activity in PAM bioreactors (both with and without coupled ZVI filter) significantly decreased. This was consistent with a sharp fall of methane productivity in those bioreactors in period 2. During period 2, PCE removals in the range 86 to 97 % were generally observed; the highest removal corresponded to PAM bioreactors along with the highest dehalogenation efficiency (94 %). Principal component analysis as well as cluster analysis confirmed the trends mentioned above, i.e., the better performance of PAM over MD, and the unexpected no effect of the ZVI side filters on PCE removal and dehalogenation efficiencies. To the best of our knowledge, this is the first report on the combined treatment ZVI-biological of a water polluted with PCE, where the biological operation relied on simultaneous electron acceptors.

Enzymatic saccharification of sugar cane bagasse by continuous xylanase and cellulase production from cellulomonas flavigena PR-22.

Cellulase (CMCase) and xylanase enzyme production and saccharification of sugar cane bagasse were coupled into two stages and named enzyme production and sugar cane bagasse saccharification. The performance of Cellulomonas flavigena (Cf) PR-22 cultured in a bubble column reactor (BCR) was compared to that in a stirred tank reactor (STR). Cells cultured in the BCR presented higher yields and productivity of both CMCase and xylanase activities than those grown in the STR configuration. A continuous culture with Cf PR-22 was run in the BCR using 1% alkali-pretreated sugar cane bagasse and mineral media, at dilution rates ranging from 0.04 to 0.22 1/h. The highest enzymatic productivity values were found at 0.08 1/h with 1846.4 ± 126.4 and 101.6 ± 5.6 U/L·h for xylanase and CMCase, respectively. Effluent from the BCR in steady state was transferred to an enzymatic reactor operated in fed-batch mode with an initial load of 75 g of pretreated sugar cane bagasse; saccharification was then performed in an STR at 55°C and 300 rpm for 90 h. The constant addition of fresh enzyme as well as the increase in time of contact with the substrate increased the total soluble sugar concentration 83% compared to the value obtained in a batch enzymatic reactor. This advantageous strategy may be used for industrial enzyme pretreatment and saccharification of lignocellulosic wastes to be used in bioethanol and chemicals production from lignocellulose. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:321-326, 2016.

Biodegradability of Nonionic Surfactant Used in the Remediation of Groundwaters Polluted with PCE.

The objective of this work was to evaluate the degradation of the nonionic surfactant Tween 80 by a PCE-degrading consortium anchored in bioparticles of fluidized bed bioreactors used in onsite remediation. Batch lab-scale bioreactors were set with dominant denitrifying (DN), methanogenic (M), and aerobic (Ab) metabolisms. Tween 80 at 100 mg/L was the sole source of carbon and energy. Denitrifying bioreactors had the highest surfactant removal (70%). Tween removals in M and Ab bioreactors were 53 and 37%, respectively. Removals of organic matter (COD) closely followed the efficiencies reported for Tween. This strongly suggested that degradation of Tween 80 occurred. Positive consequences of Tween degradation in remediation are first, the surfactant will not become an environmental/health liability by remaining as a recalcitrant or toxic substance in aquifers or in treated effluents; and second, savings on aeration could be achieved by conducting Tween 80 degradation in anaerobic conditions, either DN or M.

Long-term response on growth, antioxidant enzymes, and secondary metabolites in salicylic acid pre-treated Uncaria tomentosa microplants.

OBJECTIVE: To obtain micro propagated Uncaria tomentosa plantlets with enhanced secondary metabolites production, long-term responses to salicylic acid (SA) pre-treatments at 1 and 100 µM were evaluated after propagation of the plantlets in a SA-free medium. RESULTS: SA pre-treatments of single node cuttings OF U. tomentosa produced long-term responses in microplants grown for 75 days in a SA-free medium. Reduction in survival rate, root formation, and stem elongation were observed only with 100 µM SA pre-treatments with respect to the control (0 + DMSO).Both pre-treatments enhanced H2O2 and inhibited superoxide dismutase and catalase activities, while guaiacol peroxidase was increased only with 1 µM SA. Also, both pre-treatments increased total monoterpenoid oxindole alkaloids by ca. 55 % (16.5 mg g(-1) DW), including isopteropodine, speciophylline, mitraphylline, isomitraphylline, rhynchopylline, and isorhynchopylline; and flavonoids by ca. 21 % (914 µg g(-1) DW), whereas phenolic compounds were increased 80 % (599 µg g(-1) DW) at 1 µM and 8.2 % (359 µg g(-1) DW) at 100 µM SA. CONCLUSION: Pre-treatment with 1 µM SA of U.tomentosa microplants preserved the survival rate and increased oxindole alkaloids, flavonoids, and phenolic compounds in correlation with H2O2 and peroxidase activity enhancements, offering biotechnological advantages over non-treated microplants.

Steady-state inhibition model for the biodegradation of sulfonated amines in a packed bed reactor.

Aromatic amines are important industrial products having in their molecular structure one or more aromatic rings. These are used as precursors for the synthesis of dyes, adhesives, pesticides, rubber, fertilizers and surfactants. The aromatic amines are common constituents of industrial effluents, generated mostly by the degradation of azo dyes. Several of them are a threat to human health because they can by toxic, allergenic, mutagenic or carcinogenic. The most common are benzenesulfonic amines, such as 4-ABS (4-aminobenzene sulfonic acid) and naphthalene sulfonic amines, such as 4-ANS (4-amino naphthalene sulfonic acid). Sometimes, the mixtures of toxic compounds are more toxic or inhibitory than the individual compounds, even for microorganisms capable of degrading them. Therefore, the aim of this study was to evaluate the degradation of the mixture 4-ANS plus 4-ABS by a bacterial community immobilized in fragments of volcanic stone, using a packed bed continuous reactor. In this reactor, the amines loading rates were varied from 5.5 up to 69 mg L(-1) h(-1). The removal of the amines was determined by high-performance liquid chromatography and chemical oxygen demand. With this information, we have studied the substrate inhibition of the removal rate of the aromatic amines during the degradation of the mixture of sulfonated aromatic amines by the immobilized microorganisms. Experimental results were fitted to parabolic, hyperbolic and linear inhibition models. The model that best characterizes the inhibition of the specific degradation rate in the biofilm reactor was a parabolic model with values of RXM=58.15±7.95 mg (10(9) cells h)(-1), Ks=0.73±0.31 mg L(-1), Sm=89.14±5.43 mg L(-1) and the exponent m=5. From the microbial community obtained, six cultivable bacterial strains were isolated and identified by sequencing their 16S rDNA genes. The strains belong to the genera Variovorax, Pseudomonas, Bacillus, Arthrobacter, Nocardioides and Microbacterium. This microbial consortium could use the mixture of aromatic amines as sources of carbon, nitrogen, energy and sulfur.

Biowaste biorefinery in Europe: opportunities and research & development needs.

This review aims to explore the needs and opportunities of research & development in the field of biowaste biorefinery in Europe. Modern industry in recent years is giving its close attention on organic waste as a new precious bioresource. Specific biowaste valorisation pathways are focusing on food processing waste, being food sector the first manufacture in Europe. Anyway they need to be further tested and validated and then transferred at the larger scale. In particular, they also need to become integrated, combining biomass pretreatments and recovery of biogenic chemicals with bioconversion processes in order to obtain a large class of chemicals. This will help to (a) use the whole biowaste, by avoiding producing residues and providing to the approach the required environmental sustainability, and (b) producing different biobased products that enter different markets, to get the possible economical sustainability of the whole biorefinery. However, the costs of the developed integrated processes might be high, mostly for the fact that the industry dealing with such issues is still underdeveloped and therefore dominated by high processing costs. Such costs can be significantly reduced by intensifying research & development on process integration and intensification. The low or no cost of starting material along with the environmental benefits coming from the concomitant biowaste disposal would offset the high capital costs for initiating such a biorefinery. As long as the oil prices tend to increase (and they will) this strategy will become even more attractive.

Biohydrogen, biomethane and bioelectricity as crucial components of biorefinery of organic wastes: a review.

Biohydrogen is a sustainable form of energy as it can be produced from organic waste through fermentation processes involving dark fermentation and photofermentation. Very often biohydrogen is included as a part of biorefinery approaches, which reclaim organic wastes that are abundant sources of renewable and low cost substrate that can be efficiently fermented by microorganisms. The aim of this work was to critically assess selected bioenergy alternatives from organic solid waste, such as biohydrogen and bioelectricity, to evaluate their relative advantages and disadvantages in the context of biorefineries, and finally to indicate the trends for future research and development. Biorefining is the sustainable processing of biomass into a spectrum of marketable products, which means: energy, materials, chemicals, food and feed. Dark fermentation of organic wastes could be the beach-head of complete biorefineries that generate biohydrogen as a first step and could significantly influence the future of solid waste management. Series systems show a better efficiency than one-stage process regarding substrate conversion to hydrogen and bioenergy. The dark fermentation also produces fermented by-products (fatty acids and solvents), so there is an opportunity for further combining with other processes that yield more bioenergy. Photoheterotrophic fermentation is one of them: photosynthetic heterotrophs, such as non-sulfur purple bacteria, can thrive on the simple organic substances produced in dark fermentation and light, to give more H2. Effluents from photoheterotrophic fermentation and digestates can be processed in microbial fuel cells for bioelectricity production and methanogenic digestion for methane generation, thus integrating a diverse block of bioenergies. Several digestates from bioenergies could be used for bioproducts generation, such as cellulolytic enzymes and saccharification processes, leading to ethanol fermentation (another bioenergy), thus completing the inverse cascade. Finally, biohydrogen, biomethane and bioelectricity could contribute to significant improvements for solid organic waste management in agricultural regions, as well as in urban areas.

Extracellular expression of glucose inhibition-resistant Cellulomonas flavigena PN-120 ß-glucosidase by a diploid strain of Saccharomyces cerevisiae.

The catalytic fraction of the Cellulomonas flavigena PN-120 oligomeric ß-glucosidase (BGLA) was expressed both intra- and extracellularly in a recombinant diploid of Saccharomyces cerevisiae, under limited nutrient conditions. The recombinant enzyme (BGLA¹5) expressed in the supernatant of a rich medium showed 582 IU/L and 99.4 IU/g dry cell, with p-nitrophenyl-ß-D-glucopyranoside as substrate. BGLA¹5 displayed activity against cello-oligosaccharides with 2-5 glucose monomers, demonstrating that the protein is not specific for cellobiose and that the oligomeric structure is not essential for ß-D-1,4-bond hydrolysis. Native ß-glucosidase is inhibited almost completely at 160 mM glucose, thus limiting cellobiose hydrolysis. At 200 mM glucose concentration, BGLA¹5 retained more than 50 % of its maximal activity, and even at 500 mM glucose concentration, more than 30 % of its activity was preserved. Due to these characteristics of BGLA¹5 activity, recombinant S. cerevisiae is able to utilize cellulosic materials (cello-oligosaccharides) to produce bioethanol.

Use of organic waste for the production of added-value holocellulases with Cellulomonas flavigena PR-22 and Trichoderma reesei MCG 80.

We evaluated the production of holocellulases from the cellulolytic microorganisms Cellulomonas flavigena PR-22 and Trichoderma reesei MCG 80 using as substrates the organic fraction of municipal solid waste (OFMSW) and digestates from a hydrogenogenic-methanogenic bioenergy production process. The first set of experiments (E1) used the mutant actinobacteria C. flavigena PR-22 whereas another set (E2) used the mutant filamentous fungi T. reesei MCG 80. In E1 with OFMSW as substrate, xylanolytic activities ranged from 1800 to 3900 international units g(holocellulose)(-1) (IU g(hol)(-1)), whereas the cellulolytic activities ranged from 220 to 420 IU g(hol)(-1). The variation of agitation speed did not have a significant effect on enzyme activity, whereas the increase of substrate concentration had a significant negative effect on both xylanolytic and cellulolytic activities on a holocellulose feed basis. Regarding E2, the OFMSW was evaluated at 1, 2 and 3 % volatile solids (VS). At 2 % VS the best filter paper activities were 1200 filter paper units (FPU) l(-1); however, in a holocellulase basis the best result was 67 FPU g(hol)(-1), corresponding to 1 % VS. Next, OFMSW was compared with OFMSW supplemented with lactose, digested solids from hydrogenogenic fermentation (D1) and digested solids from a two-stage process (D2). Against expectations, no positive effect was found in OFMSW due to lactose. The best enzymatic titres were in the order D1 > OFMSW ≈ OFMSW + lactose > D2. The use of digestates from hydrogenogenic fermentation for enzyme production holds promise for waste management. It promotes energy and added-value bioproduct generation-a green alternative to common practice of management and disposal of organic wastes.