Sustainable production of bioemulsifiers, a critical overview from microorganisms to promising applications.

Microbial bioemulsifiers are molecules of amphiphilic nature and high molecular weight that are efficient in emulsifying two immiscible phases such as water and oil. These molecules are less effective in reducing surface tension and are synthesized by bacteria, yeast and filamentous fungi. Unlike synthetic emulsifiers, microbial bioemulsifiers have unique advantages such as biocompatibility, non-toxicity, biodegradability, efficiency at low concentrations and high selectivity under different conditions of pH, temperature and salinity. The adoption of microbial bioemulsifiers as alternatives to their synthetic counterparts has been growing in ongoing research. This article analyzes the production of microbial-based emulsifiers, the raw materials and fermentation processes used, as well as the scale-up and commercial applications of some of these biomolecules. The current trend of incorporating natural compounds into industrial formulations indicates that the search for new bioemulsifiers will continue to increase, with emphasis on performance improvement and economically viable processes.

Production of ß-glucan exopolysaccharide lasiodiplodan by Lasiodiplodia theobromae CCT 3966 from corn bran acid hydrolysate.

The potential utilization of corn bran acid hydrolysate (CBAH) was evaluated as an inexpensive feedstock for the production of a rich carbohydrate and protein medium for lasiodiplodan (LAS) production using the filamentous fungus Lasiodiplodia theobromae CCT 3966. Experiments were performed according to a 22 CCRD experimental design aiming to evaluate the influence of agitation speed (rpm) and temperature (°C) over the production of total cell biomass (TCB) and LAS concentration released to the medium (LAS-M), adhered to biomass (LAS-C), and total (LAS-T). Under the selected conditions (temperature of 28°C and agitation of 200 rpm), 8.73 g·L-1 of LAS-T and 4.47 g·L-1 of TCB were obtained. Recovery of LAS-C with hot water was shown as an alternative to increase the production concentration, although it might require further purification steps. CBAH potential for substitution of synthetic media was demonstrated, indicating that it is an adequate raw material containing all necessary nutrients for LAS production.Key points• Corn bran acid hydrolysate is presented as a suitable substrate for ß-glucan production.• Lasiodiplodia theobromae CCT 3966 have the potential for the industrial ß-glucan production.• Simple recovering of biomass-adhered lasiodiplodan by hot water extraction.

Xylitol bioproduction: state-of-the-art, industrial paradigm shift, and opportunities for integrated biorefineries.

Recent advances in biomass conversion technologies have shown a promising future toward fermentation during xylitol production. Xylitol is one of the top 12 renewable added-value chemicals that can be obtained from biomass according to US Department of Energy (USDOE). Currently, xylitol accounts for approximately US$823.6 million of annual sales in the market, and this amount is expected to reach US$1.37 billion by 2025. This high demand has been achieved owing to the chemical conversion of hemicellulosic hydrolysates from different lignocellulosic biomasses, which is a costly and non-ecofriendly process. Xylose-rich hemicellulosic hydrolysates are the major raw materials for xylitol production through either chemical or biotechnological routes. Economic production of a clean hemicellulosic hydrolysate is one of the major bottlenecks for xylitol production on the commercial scale. Advancements in biotechnology, such as the isolation of novel microorganisms, genetic manipulation of xylose metabolizing strains, and modifications in the fermentation process, can enhance the economic feasibility of xylitol production on the large scale. Furthermore, xylitol production in integrated biorefineries can be even more economic, given the readily available raw materials and the co-use of steam, electricity, and water, among others. Exploring new biotechnology techniques in integrated biorefineries would open new markets and opportunities for sustainable xylitol production to fulfill the market's growing demands for this sugar alcohol. This article is a review of the advancements reported in the whole biotechnological process for xylitol production, and involve pretreatment technologies, hemicellulosic hydrolysate preparation, xylose conversion into xylitol, and product recovery. Special attention is devoted to current metabolic engineering strategies to improve this bioprocess, as well as to the importance of xylitol production processes in biorefineries.

Hydrodynamic cavitation as a strategy to enhance the efficiency of lignocellulosic biomass pretreatment.

Hydrodynamic cavitation (HC) is a process technology with potential for application in different areas including environmental, food processing, and biofuels production. Although HC is an undesirable phenomenon for hydraulic equipment, the net energy released during this process is enough to accelerate certain chemical reactions. The application of cavitation energy to enhance the efficiency of lignocellulosic biomass pretreatment is an interesting strategy proposed for integration in biorefineries for the production of bio-based products. Moreover, the use of an HC-assisted process was demonstrated as an attractive alternative when compared to other conventional pretreatment technologies. This is not only due to high pretreatment efficiency resulting in high enzymatic digestibility of carbohydrate fraction, but also, by its high energy efficiency, simple configuration, and construction of systems, besides the possibility of using on the large scale. This paper gives an overview regarding HC technology and its potential for application on the pretreatment of lignocellulosic biomass. The parameters affecting this process and the perspectives for future developments in this area are also presented and discussed.

Biocompatibility of Brazilian native yeast-derived sophorolipids and Trichoderma harzianum as plant-growth promoting bioformulations.

The replacement of agrochemicals by biomolecules is imperative to mitigate soil contamination and inactivation of its core microbiota. Within this context, this study aimed at the interaction between a biological control agent such as Trichoderma harzianum CCT 2160 (BF-Th) and the biosurfactants (BSs) derived from the native Brazilian yeast Starmerella bombicola UFMG-CM-Y6419. Thereafter, their potential in germination of Oryza sativa L. seeds was tested. Both bioproducts were produced on site and characterized according to their chemical composition by HPLC-MS and GC-MS for BSs and SDS-PAGE gel for BF-Th. The BSs were confirmed to be sophorolipids (SLs) which is a well-studied compound with antimicrobial activity. The biocompatibility was examined by cultivating the fungus with SLs supplementation ranging from 0.1 to 2 g/L in solid and submerged fermentation. In solid state fermentation the supplementation of SLs enhanced spore production, conferring the synergy of both bioproducts. For the germination assays, bioformulations composed of SLs, BF-Th and combined (SLT) were applied in the germination of O. sativa L seeds achieving an improvement of up to 30% in morphological aspects such as root and shoot size as well as the presence of lateral roots. It was hypothesized that SLs were able to regulate phytohormones expression such as auxins and gibberellins during early stage of growth, pointing to their novel plant-growth stimulating properties. Thus, this study has pointed to the potential of hybrid bioformulations composed of biosurfactants and active endophytic fungal spores in order to augment the plant fitness and possibly the control of diseases.

Fermentative production of ribonucleotides from whey by Kluyveromyces marxianus: effect of temperature and pH.

Ribonucleotides have shown many promising applications in food and pharmaceutical industries. The aim of the present study was to produce ribonucleotides (RNA) by Kluyveromyces marxianus ATCC 8,554 utilizing cheese whey, a dairy industry waste, as a main substrate under batch fermentation conditions. The effects of temperature, pH, aeration rate, agitation and initial cellular concentration were studied simultaneously through factorial design for RNA, biomass production and lactose consumption. The maximum RNA production (28.66 mg/g of dry biomass) was observed at temperature 30°C, pH 5.0 and 1 g/l of initial cellular concentration after 2 h of fermentation. Agitation and aeration rate did not influence on RNA concentration (p > 0.05). Maximum lactose consumption (98.7%) and biomass production (6.0 g/l) was observed after 12 h of incubation. This study proves that cheese whey can be used as an adequate medium for RNA production by K. marxianus under the optimized conditions at industrial scale.

Bioconversion of sugarcane biomass into ethanol: an overview about composition, pretreatment methods, detoxification of hydrolysates, enzymatic saccharification, and ethanol fermentation.

Depleted supplies of fossil fuel, regular price hikes of gasoline, and environmental damage have necessitated the search for economic and eco-benign alternative of gasoline. Ethanol is produced from food/feed-based substrates (grains, sugars, and molasses), and its application as an energy source does not seem fit for long term due to the increasing fuel, food, feed, and other needs. These concerns have enforced to explore the alternative means of cost competitive and sustainable supply of biofuel. Sugarcane residues, sugarcane bagasse (SB), and straw (SS) could be the ideal feedstock for the second-generation (2G) ethanol production. These raw materials are rich in carbohydrates and renewable and do not compete with food/feed demands. However, the efficient bioconversion of SB/SS (efficient pretreatment technology, depolymerization of cellulose, and fermentation of released sugars) remains challenging to commercialize the cellulosic ethanol. Among the technological challenges, robust pretreatment and development of efficient bioconversion process (implicating suitable ethanol producing strains converting pentose and hexose sugars) have a key role to play. This paper aims to review the compositional profile of SB and SS, pretreatment methods of cane biomass, detoxification methods for the purification of hydrolysates, enzymatic hydrolysis, and the fermentation of released sugars for ethanol production.

Biological detoxification of different hemicellulosic hydrolysates using Issatchenkia occidentalis CCTCC M 206097 yeast.

This work had as its main objective to contribute to the development of a biological detoxification of hemicellulose hydrolysates obtained from different biomass plants using Issatchenkia occidentalis CCTCC M 206097 yeast. Tests with hemicellulosic hydrolysate of sugarcane bagasse in different concentrations were carried out to evaluate the influence of the hydrolysate concentration on the inhibitory compounds removal from the sugarcane bagasse hydrolysate, without reduction of sugar concentration. The highest reduction values of inhibitors concentration and less sugar losses were observed when the fivefold concentrated hydrolysate was treated by the evaluated yeast. In these experiments it was found that the high sugar concentrations favored lower sugar consumption by the yeast. The highest concentration reduction of syringaldehyde (66.67%), ferulic acid (73.33%), furfural (62%), and 5-HMF (85%) was observed when the concentrated hydrolysate was detoxified by using this yeast strain after 24 h of experimentation. The results obtained in this work showed the potential of the yeast Issatchenkia occidentalis CCTCC M 206097 as detoxification agent of hemicellulosic hydrolysate of different biomass plants.

Interaction of an acidic sophorolipid biosurfactant with phosphatidylcholine model membranes.

Sophorolipids (SLs) constitute a group of unique biosurfactants (BS) in the light of their outstanding properties, among which their antimicrobial activities stand out. SLs can exist mainly in an acidic and a lactonic form, both of which display inhibitory activity. Given the amphipathic nature of SLs it is feasible that these antimicrobial actions are the result of the perturbation of the physicochemical properties of targeted membranes. Thus, in this work we have carried out a biophysical study to unveil the molecular details of the interaction of an acidic SL with a model phospholipid membrane made of 1,2-dipalmitoy-sn-glycero-3-phosphocholine (DPPC). Using differential scanning calorimetry it was found that SL altered the phase behaviour of DPPC at low molar fractions, producing fluid phase immiscibility with the result of formation of biosurfactant-enriched domains within the phospholipid bilayer. Fourier-transform infrared spectroscopy showed that SL interacted with DPPC increasing ordering of the phospholipid acyl chain palisade and hydration of the lipid/water interface. Small angle X-ray scattering showed that SL did not modify bilayer thickness in the biologically relevant Lα fluid phase. SL was found to induce contents leakage in 1-palmitoy-2-oleoy-sn-glycero-3-phosphocholine (POPC) unilamellar liposomes, at sublytic concentrations below the cmc. This SL-induced membrane permeabilization at concentrations below the onset for membrane solubilization can be the result of the formation of laterally segregated domains, which might contribute to provide a molecular basis for the reported antimicrobial actions of SLs.

Utilization of sugarcane straw for production of ß-glucan biopolymer by Lasiodiplodia theobromae CCT 3966 in batch fermentation process.

ß-Glucans as emerging biopolymer are widely produced by microorganisms in fermentation processes using commercial sugars which make process non-economic. Lignocellulosic substances are inexpensive carbon sources, which could be exploited for sustainable production of ß-glucans. In this study, a lignocellulosic material, namely sugarcane straw (SCS) was utilized for the production of extracellular ß-glucan by Lasiodiplodia theobromae CCT3966. SCS was subjected to acid and subsequent alkaline pretreatment, followed by enzymatic saccharification using cellulase enzyme. Quantity of 48.65 g/L glucose was released after enzymatic hydrolysis. ß-Glucan production was performed by cultivation of fungal strain in SCS hydrolysate at 28 °C and initial culture pH 7. Highest ß-glucan yield and productivity of 0.047 gg-1 and 0.014 gL-1h-1, respectively was obtained at 72 h fermentation time. Kinetic study of ß-glucan production revealed experimental biosynthesis of ß-glucan from SCS hydrolysate followed the trend generated by Logistic and Luedeking-Piret models. Chemical structure of biopolymer produced showed ß-glucan constitution.

Hydrodynamic cavitation-assisted continuous pre-treatment of sugarcane bagasse for ethanol production: Effects of geometric parameters of the cavitation device.

For biotechnological conversion of lignocellulosic biomass, a pre-treatment step is required before enzymatic hydrolysis of carbohydrate fractions of the material, which is required to produce fermentable sugars for generation of ethanol or other products in a biorefinery. The most of the reported pre-treatment technologies are in batch operation mode, presenting some disadvantages as dead times in the process. In this context, hydrodynamic cavitation (HC)-assisted alkaline hydrogen peroxide (AHP) pre-treatment in continuous process was proposed for pre-treatment of sugarcane bagasse (SCB). The system was designed with a main reactor containing two devices to generate cavitation by passing liquid medium through orifice plates. For SCB pretreated in continuous process, 52.79, 34.31, 22.13 and 15.81 g of total reducing sugars (TRS) per 100 g of SCB were released in samples pretreated using orifice plates with a number of holes of 24 (d = 0.45 mm), 16 (d = 0.65 mm), 12 (d = 0.8 mm) and 8 (d = 1 mm), respectively. Computational Fluid Dynamics (CFD) tools showed that 0.94 of vapor phase volume fraction and 0.19 of cavitation number were achieved at 31 m/s of throat velocity and upstream pressure of 350,000 Pa. By using pretreated SCB, 28.44 g of ethanol/L (84.31% of yield respect to theoretical value) was produced by immobilized Scheffersomyces stipitis NRRL-Y7124 in a simultaneous hydrolysis and fermentation process at high solid loading (16% S/L). Thus, HC-assisted process was proved as a promising technology for valorization of lignocellulosic biomass.

A New Approach for the Production of Selenium-Enriched and Probiotic Yeast Biomass from Agro-Industrial by-Products in a Stirred-Tank Bioreactor.

The production of biomolecules using agro-industrial by-products as feedstock is a growing trend worldwide. Selenium (Se) is a trace element essential for health, and the Se-enrichment of yeast biomass can enhance its benefits. This study investigated the feasibility of the production of Saccharomyces cerevisiae Se-enriched biomass using a medium composed of corn bran and soybean bran acid hydrolysates as carbon and nitrogen sources in a stirred-tank reactor. After hydrolysis, hydrolysates presented complex composition and high concentrations of sugars, proteins, and minerals. The use of a stirred-tank bioreactor leads to the production of 9 g/L S. cerevisiae biomass enriched with 236.93 µg/g Se, and 99% cell viability. Likewise, the combination of sugarcane molasses and soybean bran hydrolysate was effective for cell growth of a probiotic strain of S. cerevisiae with a 24.08% ß-glucan content. The results demonstrated that starchy acid hydrolysates are low-cost and efficient substrates for the production of yeast biomass and derivate products and may contribute to further studies for a sustainable development of biorefinery technologies.

Low-pressure homogenization of tomato juice using hydrodynamic cavitation technology: Effects on physical properties and stability of bioactive compounds.

Hydrodynamic cavitation (HC) technology, offers benefits in many processes, specifically due to its low operational cost and simple configuration. In the present work, physical properties, microbial quality and stability of bioactive compounds of HC-treated tomato juice were evaluated considering different processing conditions. Significant effects of the induced cavitation such as a reduction in particle size and an increase in the apparent viscosity were observed in the HC-treated samples. Moreover, the HC-treated tomato juice showed higher stability without sedimentation for 14 days compared to a sedimentation index of 68% observed in samples before treatment and control experiment. Lycopene and phenolic contents in tomato juice were not altered even after HC-treatment. Finally, the microbiological quality was also improved, mainly for the samples treated above 55 °C. Therefore, this technology can be envisaged as a promising strategy for food processing to enhancing desirable properties in juices.

Overcoming challenges in lignocellulosic biomass pretreatment for second-generation (2G) sugar production: emerging role of nano, biotechnological and promising approaches.

Production of green chemicals and biofuels in biorefineries is the potential alternative for petrochemicals and gasoline in transitioning of petro-economy into bioeconomy. However, an efficient biomass pretreatment process must be considered for the successful deployment of biorefineries, mainly for use of lignocellulosic raw materials. However, biomass recalcitrance plays a key role in its saccharification to obtain considerable sugar which can be converted into ethanol or other biochemicals. In the last few decades, several pretreatment methods have been developed, but their feasibility at large-scale operations remains as a persistent bottleneck in biorefineries. Pretreatment methods such as hydrodynamic cavitation, ionic liquids, and supercritical fluids have shown promising results in terms of either lignin or hemicellulose removal, thus making remaining carbohydrate fraction amenable to the enzymatic hydrolysis for clean and high amount of fermentable sugar production. However, their techno-economic feasibility at industrial scale has not been yet studied in detail. Besides, nanotechnological-based technologies could play an important role in the economically viable 2G sugar production in future. Considering these facts, in the present review, we have discussed the existing promising pretreatment methods for lignocellulosic biomass and their challenges, besides this strategic role of nano and biotechnological approaches towards the viability and sustainability of biorefineries is also discussed.

The path forward for lignocellulose biorefineries: Bottlenecks, solutions, and perspective on commercialization.

Lignocellulose biorefinery encompasses process engineering and biotechnology tools for the processing of lignocellulosic biomass for the manufacturing of bio-based products (such as biofuels, bio-chemicals, biomaterials). While, lignocellulose biorefinery offers clear value proposition, success at industrial level has not been vibrant for the commercial production of renewable chemicals and fuels. This is because of high capital and operating expenditures, irregularities in biomass supply chain, technical process immaturity, and scale up challenges. As a result, commercial production of biochemicals and biofuels with right economics is still lagging behind. To hit the market place, efforts are underway by bulk and specialty chemicals producing companies like DSM (Succinic acid, Cellulosic ethanol), Dow-DuPont (1,3-Propanediol, 1,4-Butanediol), Clariant-Global bioenergies-INEOS (bio-isobutene), Braskem (Ethylene, polypropylene), Raizen, Gran-bio and POET-DSM (Cellulosic ethanol), Amyris (Farnesene), and several other potential players. This paper entails the concept of lignocellulose biorefinery, technical challenges for industrialization of renewable fuels and bulk chemicals and future directions.

A new approach for bioethanol production from sugarcane bagasse using hydrodynamic cavitation assisted-pretreatment and column reactors.

Hydrodynamic cavitation (HC) was adopted to assist alkaline-hydrogen peroxide pretreatment of sugarcane bagasse (SCB). In the following condition: 0.29 M of NaOH, 0.78% (v/v) of H2O2, 9.95 min of process time and 3 bar of inlet pressure, 95.4% of digestibility of cellulosic fraction was achieved. To take the best use of the pretreated biomass, the overall process was intensified by way of employing a packed bed flow-through column reactor and thus enabling to handle a high solid loading of 20%, thereby leading to cellulose and hemicellulose conversions to 74.7% and 75%, respectively. In the fermentation step, a bubble column reactor was introduced to maximize ethanol production from the pretreated SCB by Scheffersomyces stipitis NRRL-Y7124, resulting in 31.50 g/L of ethanol, 0.49 g/g of ethanol yield and 0.68 g/L.h of productivity. All this showed that our HC-assisted NaOH-H2O2 pretreatment strategy along with the process intensification approach might offer an option for SCB-based biorefineries.

Sugarcane bagasse hydrolysate as a potential feedstock for red pigment production by Monascus ruber.

Sugarcane bagasse (SCB) hydrolysate could be an interesting source for red pigment production by Monascus ruber Tieghem IOC 2225. The influence of different wavelength of light-emitting diode (LED) at 250 µmol.m-2.s-1 of photon flux density on red pigment production by M. ruber in glucose-based medium was evaluated. Then, SCB hydrolysate was used as carbon source under the previously selected light incidence conditions. In glucose-based medium, the highest pigment production was achieved in fermentation assisted with orange LED light (8.28 UA490nm), white light (8.26 UA490nm) and under dark condition (7.45 UA490nm). By using SCB hydrolysate-based medium, the highest red pigment production (18.71 AU490nm) was achieved under dark condition and the glucose and cellobiose present in the hydrolysate were metabolized. SCB enzymatic hydrolysate was demonstrated to be a promising carbon source for high thermal stability red pigment production (activation energy of 10.5 kcal.mol-1), turning an interesting alternative for implementation in biorefineries.

Enhanced Production of Xylitol from Poplar Wood Hydrolysates Through a Sustainable Process Using Immobilized New Strain Candida tropicalis UFMG BX 12-a.

A new strain, Candida tropicalis UFMG BX 12-a, was found to produce higher yields of xylitol on poplar wood hemicellulose hydrolysate. The hemicellulose hydrolysate liquor was detoxified using a novel method we developed, involving vacuum evaporation and solvent separation of inhibitors which made the hydrolysate free of toxins while retaining high concentrations of fermentable sugars. The effect of the detoxification method on the fermentation was also reported and compared to well-known methods reported in literature. In this study, the new strain C. tropicalis UFMG BX 12-a was used on the detoxified hydrolysate to produce xylitol. It was also compared to Candida guilliermondii FTI 20037, which has been reported to be one of the best strains for fermentative production of xylitol. To further improve the efficiency of the fermentation process, these strains were immobilized in calcium alginate beads. The yield (0.92 g g-1) and productivity (0.88 g L-1 h-1) obtained by fermenting the wood hydrolysate detoxified by our new detoxification technique using an immobilized new Candida strain were found to be higher than the values reported in literature.

Hydrodynamic cavitation as an efficient pretreatment method for lignocellulosic biomass: A parametric study.

Hydrodynamic cavitation (HC), which is a highly destructive force, was employed for pretreatment of sugarcane bagasse (SCB). The efficacy of HC was studied using response surface methodology (RSM) with determining parameters varied: inlet pressure of 1-3bar, temperature of 40-70°C, and alkaline concentration of 0.1-0.3M. At the best condition (3bar, 70°C and 0.3M NaOH), 93.05% and 94.45% of hydrolysis yield of cellulose and hemicellulose, respectively, were obtained within 30min of pretreatment time. Also, pretreatment time higher than 10min had little to do regarding to SCB composition changes using different orifice plates (16 and 27 holes, with corresponding cavitation number of 0.017 and 0.048, respectively), with higher hydrolysis yield observed at 20min of process. Therefore, HC-based approach could lead to a high yield of hydrolysis, as long as a treatment condition was right; it could be so at mild conditions and at short running time.

Ethanol production in a simultaneous saccharification and fermentation process with interconnected reactors employing hydrodynamic cavitation-pretreated sugarcane bagasse as raw material.

In this study, sugarcane bagasse (SCB) pretreated with alkali assisted hydrodynamic cavitation (HC) was investigated for simultaneous saccharification and fermentation (SSF) process for bioethanol production in interconnected column reactors using immobilized Scheffersomyces stipitis NRRL-Y7124. Initially, HC was employed for the evaluation of the reagent used in alkaline pretreatment. Alkalis (NaOH, KOH, Na2CO3, Ca(OH)2) and NaOH recycled black liquor (successive batches) were used and their pretreatment effectiveness was assessed considering the solid composition and its enzymatic digestibility. In SSF process using NaOH-HC pretreatment SCB, 62.33% of total carbohydrate fractions were hydrolyzed and 17.26g/L of ethanol production (0.48g of ethanol/g of glucose and xylose consumed) was achieved. This proposed scheme of HC-assisted NaOH pretreatment together with our interconnected column reactors showed to be an interesting new approach for biorefineries.