Evaluation of The Production of Second Generation Ethanol by Co-Culture of Saccharomyces Cerevisiae and Pachysolen Tannophilus Immobilized in Sodium Alginate.

The production of an alternative form of fuel that replaces fossil fuels has been increasingly studied due to the environmental impacts generated by its excessive use, as well as the depletion of these fossil energy sources. Ethanol obtained from the crushing of sugar cane has been used as a substitute for these fuels, mainly in the automotive area. However, alternative sources are being studied to produce the so called second generation bioethanol. This would avoid competition for food producing agricultural areas and agroindustrial waste is a great source for obtaining it. In general, these residues are not always completely reused and are disposed of inappropriately in the environment, becoming contaminants. Therefore, the use of agroindustrial waste can become a renewable source of energy, in addition to reducing environmental impacts. The objective of this work is to produce second generation bioethanol as an alternative to the one currently used, using the rice husk hydrolyzate by the consortium formed by Saccharomyces cerevisiae and Pachysolen tannophilus . For this, an acid hydrolysis was performed with 2% sulfuric acid during 10 minutes of heating in an autoclave, after which the hydrolyzate was detoxified with the use of activated carbon. The crude and detoxified hydrolysates were used as a substrate for the fermentation medium with an initial concentration of 50 mg/mL of reducing sugars. The fermentation process with the use of both yeasts in the crude hydrolyzate medium, in the detoxified medium and in a synthetic medium composed of glucose, was carried out for 24 h, 30º C, 0 rpm and pH 6.5.The best results for the ethanol production of Saccharomyces cerevisiae was the synthetic medium with 20.6 mg/mL. For the yeast Pachysolen tannophilus , its highest production was in a synthetic medium with 11.67 mg/mL. The intercropping of the two yeasts proved to be efficient with a greater ethanol production reaching 21.5 mg/mL, the hydrolyzed and detoxified media showed great potential for ethanol production both in intercropping and in monoculture

Evaluation of Crystalline Cellulose of Corn Straw through Different Pretreatments Via X-Ray Diffraction, Scanning Electron Microscopy and Infrared Spectroscopy

Waste recycling is beneficial not only for the environment but also for the economy and the society at large. Corn stalks, the most abundant crop waste, are processed with oven drying, steaming, complex enzymatic hydrolysis (cellulase, xylanase, β-glucanase and pectinase), and fermentation (by Candida utilis and Pachysolen tannophilus), were analyzed via X-ray diffraction, Scanning Electron Microscopy (SEM) and Infrared Spectroscopy (IR). The results indicated that thermophilic digestion destroyed glycosidic bonds and fibrous crystal have a significant effect on the degradation of the corn stalks, while complex enzyme hydrolysis only slightly degraded fibrous crystal. Fermentation did not significantly reduce fibrous crystal. Therefore, our research suggested that thermophilic digestion is the appropriate way to increase the saccharification rate and feed yield of corn stalks.

Biochemical Conversion of Lignocellulosic Biomass from Date Palm of Phoenix dactylifera L. into Ethanol Production

Cellulosic fibers from date palm are among the most promising lignocellulose feedstock for biorefinery purposes. The world production is between 1.9 and 2.4 million t/year. Initially, a pretreatment with dilute-sulphuric acid of these fibers was performed using a response surface methodology, with temperature and process time as factors. The aim is to produce bioethanol from young and old fibers from date palm, Phoenix dactylifera L. Optimal thermochemical pretreatment conditions for both fibers palms were 220 °C in hydrothermal conditions (without acid); in these conditions pretreated young fibers presented a maximum content in holocelluloses of 45.18% and old fibers 61.97%. Subsequently, during the enzymatic hydrolysis a maximum yield of total reducing sugars (TRS) was reached, 46.32 g/100 g for pretreated dry young fibers and 48.54 g/100 g for pretreated dry old fibers. After enzymatic saccharification, hydrolysates were fermented by Pachysolen tannophilus (ATCC 32691) to ethanol, reaching yields (YE/TRS) of 37.94 g ethanol/100 g of TRS for young fibers and 35.84 g ethanol/100 g of TRS for old fibers. Globally, considering the full process, in the fermentation of the hydrolysates, a yield (YE) of 10.64 g ethanol/100 g of dry young fibers and 10.88 g ethanol/100 g of dry old fibers was reached.

Ethanol Production from Olive Stones through Liquid Hot Water Pre-Treatment, Enzymatic Hydrolysis and Fermentation. Influence of Enzyme Loading, and Pre-Treatment Temperature and Time

Olive table industry, olive mills and olive pomace oil extraction industries annually generate huge amounts of olive stones. One of their potential applications is the production of bioethanol by fractionation of their lignocellulose constituents and subsequent fermentation of the released sugars using yeasts. In this work, we studied the influence of temperature (175–225 °C) and residence time (0–5 min) in the liquid hot-water pre-treatment of olive stones as well as the initial enzyme loading (different mixtures of cellulases, hemicellulases and β–glucosidases) in the later enzymatic hydrolysis on the release of fermentable sugars. The Chrastil’s model was applied to the d-glucose data to relate the severity of pre-treatment to enzyme diffusion through the pre-treated cellulose. Finally, the hydrolysate obtained under the most suitable conditions (225 °C and 0 min for pre-treatment; 24 CE initial enzyme concentration) was fermented into ethanol using the yeast Pachysolen tannophilus ATCC 32691. Considering the overall process, 6.4 dm3 ethanol per 100 kg olive stones were produced.

Pentose metabolism and conversion to biofuels and high-value chemicals in yeasts

ABSTRACT Pentose sugars are widespread in nature and two of them, D-xylose and L-arabinose belong to the most abundant sugars being the second and third by abundance sugars in dry plant biomass (lignocellulose) and in general on planet. Therefore, it is not surprising that metabolism and bioconversion of these pentoses attract much attention. Several different pathways of D-xylose and L-arabinose catabolism in bacteria and yeasts are known. There are even more common and really ubiquitous though not so abundant pentoses, D-ribose and 2-deoxy-D-ribose, the constituents of all living cells. Thus, ribose metabolism is example of endogenous metabolism whereas metabolism of other pentoses, including xylose and L-arabinose, represents examples of the metabolism of foreign exogenous compounds which normally are not constituents of yeast cells. As a rule, pentose degradation by the wild-type strains of microorganisms does not lead to accumulation of high amounts of valuable substances; however, productive strains have been obtained by random selection and metabolic engineering. There are numerous reviews on xylose and (less) L-arabinose metabolism and conversion to high value substances; however, they mostly are devoted to bacteria or the yeast Saccharomyces cerevisiae. This review is devoted to reviewing pentose metabolism and bioconversion mostly in non-conventional yeasts which naturally metabolize xylose, namely, Scheffersomyces stipitis, Scheffersomyces shehatae, Pachysolen tannophilus, Spathaspora passalidarum, Kluyveromyces marxianus, Ogataea polymorpha, Candida intermedia, Candida tenuis, Meyerozyma guilliermondii, Komagataella phaffii, Yarrowia lipolytica. Pentose metabolism in the recombinant strains of S. cerevisiae is also considered for comparison. This review discusses recent developments in studying pentose metabolism and bioconversion to biofuels and high-value chemicals in several species of The available data on ribose, xylose, L-arabinose transport, metabolism, regulation of these processes, interaction with glucose catabolism and construction of the productive strains of high-value chemicals or pentose (ribose) itself are described. In addition, genome studies of the natural xylose metabolizing yeasts and available tools for their molecular research are reviewed. Metabolism of other pentoses (2-deoxyribose, D-arabinose, lyxose) is briefly reviewed.

Designing Efficient Processes for Sustainable Bioethanol and Bio-Hydrogen Production from Grass Lawn Waste

The effect of thermal, acid and alkali pretreatment methods on biological hydrogen (BHP) and bioethanol production (BP) from grass lawn (GL) waste was investigated, under different process schemes. BHP from the whole pretreatment slurry of GL was performed through mixed microbial cultures in simultaneous saccharification and fermentation (SSF) mode, while BP was carried out through the C5yeast Pichia stipitis, in SSF mode. From these experiments, the best pretreatment conditions were determined and the efficiencies for each process were assessed and compared, when using either the whole pretreatment slurry or the separated fractions (solid and liquid), the separate hydrolysis and fermentation (SHF) or SSF mode, and especially for BP, the use of other yeasts such as Pachysolen tannophilus or Saccharomyces cerevisiae. The experimental results showed that pretreatment with 10 gH2SO4/100 g total solids (TS) was the optimum for both BHP and BP. Separation of solid and liquid pretreated fractions led to the highest BHP (270.1 mL H2/g TS, corresponding to 3.4 MJ/kg TS) and also BP (108.8 mg ethanol/g TS, corresponding to 2.9 MJ/kg TS) yields. The latter was achieved by using P. stipitis for the fermentation of the hydrolysate and S. serevisiae for the solid fraction fermentation, at SSF.

The Process of Producing Bioethanol from Delignified Cellulose Isolated from Plants of the Miscanthus Genus

Plants of the Miscanthus genus (Miscanthus Anderss.) have a unique index of biomass production in relation to the occupied area. Miscanthus plants can be attributed to promising second-generation raw materials for the production of bioethanol and biofuel. Miscanthus plants are characterized by a high cellulose content. Herein, we report the results of a study on the obtained delignified cellulose with subsequent processing into bioethanol using microbial communities. In the course of the study, the optimal conditions for the delignification of the initial plant material for cellulose were selected. Ethanol with a high degree of conversion was successfully obtained from the isolated delignified cellulose. The article describes the pilot technological scheme for the conversion of Miscanthus plant biomass to bioethanol involving the delignification stages, followed by the conversion of the resulting cellulose into bioethanol by a consortium of microorganisms. As a result of the study, it was found that delignification using trifluoroacetic acid leads to the production of cellulose of high purity. Bioethanol with a yield of 3.1% to 3.4% in terms of the initial amount of biomass was successfully obtained by a microorganism consortium of Saccharomyces cerevisiae M Y-4242/Pachysolen tannophilus Y-3269, and Scheffersomyces stipitis Y-3264.

Acid and Enzymatic Fractionation of Olive Stones for Ethanol Production Using Pachysolen tannophilus

Olive stones are an abundant lignocellulose material in the countries of the Mediterranean basin that could be transformed to bioethanol by biochemical pathways. In this work, olive stones were subjected to fractionation by means of a high-temperature dilute-acid pretreatment followed by enzymatic hydrolysis of the pretreated solids. The hydrolysates obtained in these steps were separately subjected to fermentation with the yeast Pachysolen tannophilus ATCC 32691. Response surface methodology with two independent variables (temperature and reaction time) was applied for optimizing D-xylose production from the raw material by dilute acid pretreatment with 0.01 M sulfuric acid. The highest D-xylose yield in the liquid fraction was obtained in the pretreatment at 201 °C for 5.2 min. The inclusion of a detoxification step of the acid prehydrolysate, by vacuum distillation, allowed the fermentation of the sugars into ethanol and xylitol. The enzymatic hydrolysis of the pretreated solids was solely effective when using high enzyme loadings, thus leading to easily fermentable hydrolysates into ethanol. The mass macroscopic balances of the overall process illustrated that the amount of inoculum used in the fermentation of the acid prehydrolysates strongly affected the ethanol and xylitol yields.