Probing the polyphenolic flavonoid, morin as a highly efficacious inhibitor against amyloid(A4V) mutant SOD1 in fatal amyotrophic lateral sclerosis.

Deposition of misfolded protein aggregates in key areas of human brain is the quintessential trait of various pertinent neurodegenerative disorders including amyotrophic lateral sclerosis (ALS). Genetic point mutations in Cu/Zn superoxide dismutase (SOD1) are found to be the most important contributing factor behind familial ALS. Especially, single nucleotide polymorphism (SNP) A4V is the most nocuous since it substantially decreases life expectancy of patients. Besides, the use of naturally occurring polyphenolic flavonoids is profoundly being advocated for palliating amyloidogenic behavior of proteopathic proteins. In the present analysis, through proficient computational tools, we have attempted to ascertain a pharmacodynamically promising flavonoid compound that effectively curbs the pathogenic behavior of A4V SOD1 mutant. Initial screening of flavonoids that exhibit potency against amyloids identified morin, myricetin and epigallocatechin gallate as promising leads. Further, with the help of feasible and yet adept protein-ligand interaction studies and stalwart molecular simulation analyses, we were able to observe that aforementioned flavonoids were able to considerably divert mutant A4V SOD1 from its distinct pathogenic behavior. Among which, morin showed the most curative potential against A4V SOD1. Therefore, morin holds a great therapeutic potential in contriving highly efficacious inhibitors in mitigating fatal and insuperable ALS.

Computer aided therapeutic tripeptide design, in alleviating the pathogenic proclivities of nocuous α-synuclein fibrils.

Parkinson's disorder (PD) exacerbates neuronal degeneration of motor nerves, thereby effectuating uncoordinated movements and tremors. Aberrant alpha-synuclein (α-syn) is culpable of triggering PD, wherein cytotoxic amyloid aggregates of α-syn get deposited in motor neurons to instigate neuro-degeneration. Amyloid aggregates, typically rich in beta sheets are cardinal targets to mitigate their neurotoxic effects. In this analysis, owing to their interaction specificity, we formulated an efficacious tripeptide out of the aggregation-prone region of α-syn protein. With the help of a proficient computational pipeline, systematic peptide shortening and an adept molecular simulation platform, we formulated a tripeptide, VAV from α-syn structure based hexapeptide KISVRV. Indeed, the VAV tripeptide was able to effectively mitigate the α-syn amyloid fibrils' dynamic rate of beta-sheet formation. Additional trajectory analyses of the VAV- α-syn complex indicated that, upon its dynamic interaction, VAV efficiently altered the distinct pathogenic structural dynamics of α-syn, further advocating its potential in alleviating aberrant α-syn's amyloidogenic proclivities. Consistent findings from various computational analyses have led us to surmise that VAV could potentially re-alter the pathogenic conformational orientation of α-syn, essential to mitigate its cytotoxicity. Hence, VAV tripeptide could be an efficacious therapeutic candidate to efficiently ameliorate aberrant α-syn amyloid mediated neurotoxicity, eventually attenuating the nocuous effects of PD.Communicated by Ramaswamy H. Sarma.

Defensin-based therapeutic peptide design in attenuating V30M TTR-induced Familial Amyloid Polyneuropathy.

In the present study, we aimed to formulate an effective therapeutic candidate against V30M mutant transthyretin (TTR) protein to hinder its pathogenic misfolding. Nicotiana alata Defensin 1 (NaD1) Antimicrobial Peptide (AMP) was availed due to its tendency to aggregate, which may compete for aggregation-prone regions of pathogenic TTR protein. Based on NaD1's potential to bind to V30M TTR, we proposed NaD1-derived tetra peptides: CKTE and SKIL to be initial therapeutic candidates. Based on their association with mutant TTR protein, CKTE tetra peptide showed considerable interaction and curative potential as compared to SKIL tetra peptide. Further analyses from discrete molecular dynamics simulation corroborate CKTE tetra peptide's effectiveness as a 'beta-sheet breaker' against V30M TTR. Various post-simulation trajectory analyses suggested that CKTE tetra peptide alters the structural dynamics of pathogenic V30M TTR protein, thereby potentially attenuating its beta-sheets and impeding its aggregation. Normal mode analysis simulation corroborated that V30M TTR conformation is altered upon its interaction with CKTE peptide. Moreover, simulated thermal denaturation findings suggested that CKTE-V30M TTR complex is more susceptible to simulated denaturation, relative to pathogenic V30M TTR; further substantiating CKTE peptide's potential to alter V30M TTR's pathogenic conformation. Moreover, the residual frustration analysis augmented CKTE tetra peptide's proclivity in reorienting the conformation of V30M TTR. Therefore, we predicted that the tetra peptide, CKTE could be a promising therapeutic candidate in mitigating the amyloidogenic detrimental effects of V30M TTR-mediated familial amyloid polyneuropathy (FAP). Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03646-4.

The realm of cellulases in biorefinery development.

Geopolitical concerns (unstable supply of gasoline, environmental pollution, and regular price hikes), economic, and employment concerns have been prompting researchers, entrepreneurs, and policy makers to focus on harnessing the potential of lignocellulosic feedstock for fuel ethanol production and its commercialization. The carbohydrate skeleton of plant cell walls needs to be depolymerised into simpler sugars for their application in fermentation reactions as a chief carbon source of suitable ethnologic strains for ethanol production. The role of cellulolytic enzymes in the degradation of structural carbohydrates of the plant cell wall into ready-to-fermentable sugar stream is inevitable. Cellulase synergistically acts upon plant cell wall polysaccharides to release glucose into the liquid media. Cellulase predominantly dominates all the plant cell wall degrading enzymes due to their vast and diverse range of applications. Apart from the major applications of cellulases such as in detergent formulations, textile desizing, and development of monogastric feed for ruminants, their role in biorefinery is truly remarkable. This is a major area where new research tools based upon fermentation based formulations, biochemistry, and system biology to expedite the structure-function relationships of cellulases including cellulosomes and new designer enzymatic cocktails are required. In the last two decades, a considerable amount of research work has been performed on cellulases and their application in biomass saccharification. However, there are still technical and economic impediments to the development of an inexpensive commercial cellulase production process. Advancements in biotechnology such as screening of microorganisms, manipulation of novel cellulase encoding traits, site-specific mutagenesis, and modifications to the fermentation process could enhance the production of cellulases. Commercially, cheaper sources of carbohydrates and modified fermentation conditions could lead to more cost-effective production of cellulases with the goal to reduce the cost of ethanol production from lignocellulosics. Implementation of integrated steps like cellulase production and cellulase mediated saccharification of biomass in conjunction with the fermentation of released sugars in ethanol in a single step so called consolidated bio-processing (CBP) is very important to reduce the cost of bioethanol. This paper aims to explore and review the important findings in cellulase biotechnology and the forward path for new cutting edge opportunities in the success of biorefineries.

Investigating the pernicious effects of heparan sulfate in serum amyloid A1 protein aggregation: a structural bioinformatics approach.

Amyloid-A mediated (AA) amyloidosis is the pathogenic byproduct of body's prolonged exposure to inflammatory conditions. It is described by the aggregation of mutated/misfolded serum amyloid A1 (SAA1) protein in various tissues and organs. Genetic polymorphism G90D is suspected to cause AA amyloidosis, although the causal mechanism remains cryptic. Recent experimental findings insinuate that heparan sulphate (HS), a glycosaminoglycans, exhibits binding with SAA1 to promote its aggregation. To foster the enhanced binding of HS, we computationally determined the pernicious modifications in G90D mutant SAA1 protein. Also, we examined the influence of HS on the dynamic conformation of mutant SAA1 that could potentially succor amyloidosis. Accordingly, the protein-ligand binding studies indicate that upon SNP G90D, SAA1 protein exhibited an augmented association with HS. Further, the simulation of HS bound mutant SAA1 complex delineates an increase in RMSD, Rg, and RMSF. Also, both RMSD and Rg evinced a fluctuating trajectory. Further, the complex showed increase of beta turn in its secondary structural composition. Additionally, the free energy landscape of mutant SAA1-HS complex posits the occurrence of multiple global minima conformers as opposed to the presence of a single global energy minima conformation in native SAA1 protein. In conclusion, the aforementioned conformational ramifications induced by HS on SAA1 could potentially be the proteopathic incendiary behind AA amyloidosis; this incendiary will need to be considered in future studies for developing effective therapeutics against AA amyloidosis.Communicated by Ramaswamy H. Sarma.

Molecular simulation probes the potency of resveratrol in regulating the toxic aggregation of mutant V30M TTR fibrils in Transthyretin mediated amyloidosis.

Transthyretin (TTR) mediated amyloidosis is a highly ruinous illness that affects various organs by aggravating the deposition of misfolded or mutated TTR protein aggregates in tissues. Hence, hindering the formation of TTR amyloid aggregates could be a key strategy in finding an effective cure towards the aggravating disorder. In this analysis, we examined the subversive nature of point mutation, V30M, in TTR that promotes amyloidogenicity using discrete molecular dynamics (DMD) simulations. Besides, we probed the association of naturally occurring polyphenols: EGCG (a proven anti TTR aggregation agent as positive control), resveratrol and curcumin in mitigating the pathogenic repercussions of mutant TTR. Results from the computational studies endorsed that the resveratrol constitutes a restorative potential to subjugate TTR mediated amyloidosis, besides EGCG. Hence, this study could be a reminiscent aspect in understanding the inhibitory role of key polyphenols against the mutant TTR aggregates, which could be an aid towards structure-based drug design in the upcoming research era on familial amyloid disorders.

Molecular simulation unravels the amyloidogenic misfolding of nascent ApoA1 protein, driven by deleterious point mutations occurring in between 170-178 hotspot region.

Protein ApoA1 is extensively studied for its role in lipid metabolism. Its seedy dark side of amyloid formulation remains relatively understudied yet. Due to genetic mutations, the protein pathologically misshapes into its amyloid form that gets accumulated in various organs, including the heart. To contrive effective therapeutics against this debilitating congenital disorder, it is imperative to comprehend the structural ramifications induced by mutations in APoA1's dynamic conformation. Till now, several point mutations have been implicated in ApoA1's amyloidosis, although only a handful has been examined considerably. Especially, the single nucleotide polymorphisms (SNPs) that occur in-between 170-178 mutation hotspot site of APoA1 needs to be investigated, since most of them are culpable of amyloid deposition in the heart. To that effect, in the present study, we have computationally quantified and studied the ApoA1's biomolecular modifications fostered by SNPs in the 170-178 mutation hotspot. Findings from discrete molecular dynamics simulation studies indicate that the SNPs have noticeably steered the ApoA1's behaviour from its native structural dynamics. Analysis of protein's secondary structural changes exhibits a considerable change upon mutations. Further, subjecting the protein structures to simulated thermal denaturation shows increased resistance to denaturation among mutants when compared to native. Further, normal mode analysis of protein's dynamic motion also shows discrepancy in its dynamic structural change upon SNP. These structural digressions induced by SNPs can very well be the biomolecular incendiary that drives ApoA1 into its amyloidogenesis. And, understanding these structural modifications initiates a better understanding of SNP's amyloidogenic pathology on APoA1.Communicated by Ramaswamy H. Sarma.

Probing the competitive inhibitor efficacy of frog-skin alpha helical AMPs identified against ACE2 binding to SARS-CoV-2 S1 spike protein as therapeutic scaffold to prevent COVID-19.

In COVID-19 infection, the SARS-CoV-2 spike protein S1 interacts to the ACE2 receptor of human host, instigating the viral infection. To examine the competitive inhibitor efficacy of broad spectrum alpha helical AMPs extracted from frog skin, a comparative study of intermolecular interactions between viral S1 and AMPs was performed relative to S1-ACE2p interactions. The ACE2 binding region with S1 was extracted as ACE2p from the complex for ease of computation. Surprisingly, the Spike-Dermaseptin-S9 complex had more intermolecular interactions than the other peptide complexes and importantly, the S1-ACE2p complex. We observed how atomic displacements in docked complexes impacted structural integrity of a receptor-binding domain in S1 through conformational sampling analysis. Notably, this geometry-based sampling approach confers the robust interactions that endure in S1-Dermaseptin-S9 complex, demonstrating its conformational transition. Additionally, QM calculations revealed that the global hardness to resist chemical perturbations was found more in Dermaseptin-S9 compared to ACE2p. Moreover, the conventional MD through PCA and the torsional angle analyses indicated that Dermaseptin-S9 altered the conformations of S1 considerably. Our analysis further revealed the high structural stability of S1-Dermaseptin-S9 complex and particularly, the trajectory analysis of the secondary structural elements established the alpha helical conformations to be retained in S1-Dermaseptin-S9 complex, as substantiated by SMD results. In conclusion, the functional dynamics proved to be significant for viral Spike S1 and Dermaseptin-S9 peptide when compared to ACE2p complex. Hence, Dermaseptin-S9 peptide inhibitor could be a strong candidate for therapeutic scaffold to prevent infection of SARS-CoV-2.

Alpha-Synuclein Aggregation in Parkinson's Disease.

Parkinson's disease (PD), a neurodegenerative disorder characterized by distinct aging-independent loss of dopaminergic neurons in substantia nigra pars compacta (SNpc) region urging toward neuronal loss. Over the decade, various key findings from clinical perspective to molecular pathogenesis have aided in understanding the genetics with assorted genes related with PD. Subsequently, several pathways have been incriminated in the pathogenesis of PD, involving mitochondrial dysfunction, protein aggregation, and misfolding. On the other hand, the sporadic form of PD cases is found with no genetic linkage, which still remain an unanswered question? The exertion in ascertaining vulnerability factors in PD considering the genetic factors are to be further dissevered in the forthcoming decades with advancement in research studies. One of the major proponents behind the prognosis of PD is the pathogenic transmutation of aberrant alpha-synuclein protein into amyloid fibrillar structures, which actuates neurodegeneration. Alpha-synuclein, transcribed by SNCA gene is a neuroprotein found predominantly in brain. It is implicated in the modulation of synaptic vesicle transport and eventual release of neurotransmitters. Due to genetic mutations and other elusive factors, the alpha-synuclein misfolds into its amyloid form. Therefore, this review aims in briefing the molecular understanding of the alpha-synuclein associated with PD.

Decoding Conformational Imprint of Convoluted Molecular Interactions Between Prenylflavonoids and Aggregated Amyloid-Beta42 Peptide Causing Alzheimer's Disease.

Protein misfolding occurs due to the loss of native protein structure and adopts an abnormal structure, wherein the misfolded proteins accumulate and form aggregates, which result in the formation of amyloid fibrils that are associated with neurodegenerative diseases. Amyloid beta (Aß42) aggregation or amyloidosis is contemplated as a unique hallmark characteristic of Alzheimer's disease (AD). Due to aberrant accrual and aggregation of Aß42 in extracellular space, the formation of senile plaques is found in AD patients. These senile plaques occur usually in the cognitive and memory region of the brain, enfeebles neurodegeneration, hinders the signaling between synapse, and disrupts neuronal functioning. In recent years, herbal compounds are identified and characterized for their potential as Aß42 inhibitors. Thus, understanding their structure and molecular mechanics can provide an incredible finding in AD therapeutics. To describe the structure-based molecular studies in the rational designing of drugs against amyloid fibrils, we examined various herbal compounds that belong to prenylflavonoids. The present study characterizes the trends we identified at molecular docking studies and dynamics simulation where we observed stronger binding orientation of bavachalcone, bavachin, and neobavaisoflavone with the amyloid-beta (Aß42) fibril structure. Hence, we could postulate that these herbal compounds could be potential inhibitors of Aß42 fibrils; these anti-aggregation agents need to be considered in treating AD.

Applying the recent clinical trials on primary open angle glaucoma: the developing world perspective.

Recent clinical trials have provided scientific guidelines for the treatment of ocular hypertension and primary open angle glaucoma. The developing world need to apply these trials in a sensible and cost effective manner. The number needed to treat (NNT) attempts to tailor treatment to the individual patient. The NNT for the average ocular hypertensive is 20. Those with intraocular pressure > or =26 mm Hg have an NNT of 6. Restricting treatment to those with lower central corneal thickness and or high cup disc ratios can further lower NNT and make treatment more cost effective. The NNT for the average patient with early POAG is 5. Targeting those at higher risk for progression, (bilateral POAG, higher IOP and or pseudo-exfoliation) can further reduce NNT. As far as the modality of treatment is concerned, provided quality can be ensured, collaborative initial glaucoma treatment study (CIGTS) could be interpreted to justify primary surgery in the developing world context. Population attributable risk percentage (PAR), a measure that reflects the public health importance of a disease was used to extrapolate results to the overall population. Ocular hypertension has an "effective" PAR of 8.5%, a value not considered high enough to warrant public health intervention. POAG had an "effective" PAR of 16%, perhaps high enough to be considered a public health problem and justify inclusion as a target disease in the Vision 2020 program. However the logistics and opportunity costs of diagnosis and treatment would probably prevent inclusion of POAG in public health budgets of most developing countries.

Ultrasonic evaluation of retinoblastoma.

Seventeen children presenting with leucocoria were subjected to complete investigations. Careful A and B mode ultrasonography allowed us to make the correct diagnosis of retinoblastoma in 11 eyes of 9 patients, which was confirmed by histopathology or by response to radiotherapy. The diagnosis of retinoblastoma was not missed ultrasonographically in any of the eyes studied. The "V/W" configuration and absence of vascular pulsations on A mode; the "mixed" pattern and absence of choroidal excavation and orbital shadowing on B mode were found to be essential echographic features in characterizing the tumor and in differentiating it from other conditions including vitreous hemorrhage. The ultrasonographic features of early extraocular extension of retinoblastoma are discussed. Careful ultrasonic evaluation of leucocoria would seem to be invaluable in obviating the possibility of misdiagnosis in such problematic cases.

Humphrey visual field and frequency doubling perimetry in the diagnosis of early glaucoma.

PURPOSE: To compare Humphrey Visual Field Analyzer (HVF) and Frequency Doubling Perimetry (FDP) testing in the diagnosis of early glaucoma. METHODS: We performed HVF (24-2 standard full threshold) and FDP (N-30) evaluations in 34 consecutive patients with early primary glaucoma and 96 normal subjects. Early glaucoma was defined on the basis of disc changes of glaucoma; the mean deviation on white-on-white perimetry had to be no worse than 6 decibels. Glaucomatous optic neuropathy was defined as a combination of cup-disc asymmetry of more than 0.2, notching, excavation, thinning or pallor of superior or inferior neuroretinal rims, retinal nerve fibre layer defects of the wedge or diffuse type and neuroretinal rim haemorrhage. Both the glaucoma patients and normal subjects had vision better than 6/9 with correction. They had no media opacities other than early nuclear sclerosis and no fundus pathology. Further, normal subjects were free of systemic diseases known to affect the retina or optic nerve. The sensitivity and specificity of HVF and FDP were calculated. RESULTS: There were 44 eligible eyes among the 34 subjects. The glaucomatous disc findings included notch (n=8), pallor (n=21), thinning (n=23) and haemorrhage (n=1) of the neuroretinal rim. The sensitivity and the specificity of the HVF were 52.3% and 57.3% respectively. The sensitivity and the specificity of FDP were 65.9% and 61.5% respectively. CONCLUSION: The difference between FDP and HVF in the diagnosis of early glaucoma is not marked.

The role of artificial drainage devices in glaucoma surgery.

The use of artificial drainage devices (ADDs) or "setons" in glaucoma surgery is generally restricted to patients with refractory glaucoma at high risk for failure from conventional filtration surgery. ADDs, both valved and nonvalved are currently available in this country. Recently, some of these devices have been propogated as primary treatment even for primary glaucomas. This article examines the role of ADDs in the modern management of the glaucomas. Specific indications for ADDs and methods to reduce the complication of overfiltration are discussed. The use of antimitotics, such as 5-fluorouracil or mitomycin, with traditional filtration has decreased the indications for ADDs. The literature and our experience confirm that currently there is no role for use of ADDs as a primary procedure in most glaucomas.

Bronchial hyperresponsiveness in patients with sarcoidosis.

OBJECTIVE: To study bronchial hyperresponsiveness (BHR) in patients with sarcoidosis. METHODS: Twenty freshly diagnosed patients with sarcoidosis were evaluated. Progressively increasing concentrations of methacholine were administered using a standard five-breath dosimeter protocol, and spirometry performed after each challenge. BHR was expressed as PC20 (provocating concentration of methacholine resulting in 20% fall in forced expiratory volume in first second). All patients were nonsmokers, had no other concurrent cardiopulmonary disease, and were not on any specific treatment for sarcoidosis. RESULTS: Borderline (PC20 4.0-16.0 mg/mL), mild (PC20 1.0-4.0 mg/mL) and moderate to severe (PC20 <1.0 mg/mL) BHR was documented in three (15%), one (5%) and five (25%) patients respectively. Patients with BHR were similar to those having normal reactivity in terms of age, gender, disease duration and results of pulmonary function testing. All patients with endobronchial disease had BHR. CONCLUSION: BHR may be seen in several patients of sarcoidosis and could possibly contribute to decline in lung function.

Administration of bisphosphonate (ibandronate) impedes molar tooth movement in rabbits: A radiographic assessment.

INTRODUCTION: Bisphosphonate (Bp)-ibandronate is a pharmacological agent, exhibits antiosteoclastic or antiresorptive activity and used to treat osteolytic or osteopenic disorders. BP-ibandronate may also interfere during orthodontic tooth movement. The aim of this study was to examine the influence of locally administered Bp-ibandronate on experimental tooth movement in rabbits. MATERIALS AND METHODS: Twenty rabbits were divided into two groups- "10" served as Group-1 (control) and other "10" as Group-2 (experimental). Both groups received nickel-titanium closed coil springs with 100 g force between mandibular molar and incisors. Group-1 animals received 1 ml normal saline and Group-2 animals received ibandronate solution (0.3 mg/kg body weight) locally, mesial to the mandibular molar on the 1(st), 7(th), and 14(th) day of the experiment. A total of "40" lateral cephalograms were taken from both groups on the 1(st) and 21(st) day using a digital X-ray unit (Siemens X-ray systems, 300 mA Pleomophos analog, 2008, Germany). Individually, each animal's radiograph was traced manually and superimposed. The molar tooth movement was measured with the help of a standard metric scale. RESULTS: The Student's t-test has been done to compare the mean values of Group-1 (4.650 ± 0.363) and Group-2 (2.030 ± 0.291) and the difference was statistically significant (P < 0.001). CONCLUSION: The retarded molar tooth movement was noticed in local drug administered rabbits, which could be beneficial in orthodontics to control the undesired tooth movement.

Molecular characterization of group B rotavirus circulating in pigs from India: identification of a strain bearing a novel VP7 genotype, G21.

The occurrence of group B rotavirus (RVB) infections in pigs has been reported from different parts of world. However, such infection in the pig population maintained in Indian farms has not been investigated as yet. A total of 187 faecal specimens were collected from pigs reared in different pig farms/pigsties located in western and northern regions of India and tested for the presence of porcine RVB by amplification of the NSP2 gene using conventional RT-PCR. Nine specimens (4.8%) were shown to contain RVB RNA. N2 and N4 genotypes of NSP2 gene were detected in three and six RVB strains respectively. VP7 (G-type) and NSP5 (H-type) genes of selected six RVB strains were characterized to identify the genotypes. Multiple G (G7, G19 and G20) and H (H4 and H5) genotypes detected in the RVB strains indicated circulation of heterogeneous population of RVB strains in pigs of India. Additionally, one strain was proposed to belong to a novel RVB genotype designated as G21 on account of <80% identity of VP7 gene sequence with its counterpart in RVB strains from 20 established genotypes. Deduced amino acid sequence of VP7 gene also displayed the presence of seven unique substitutions in the strain. The study reports for the first time the occurrence of RVB infections in Indian pig herds and provides important epidemiological data useful for better understanding of ecology and evolution of porcine RVBs.

Bioconversion of novel substrate Saccharum spontaneum, a weedy material, into ethanol by Pichia stipitis NCIM3498.

Rising fuel prices and environmental issues have paved the way for the exploration of cellulosic ethanol. However, challenges involving substrate hydrolysis and cost-effectiveness still limit the efficient bioconversion and utilization of cellulosic ethanol. We aimed to evaluate a cheaper and abundantly available wild sugarcane variety, Saccharum spontaneum, as the raw substrate for bioconversion of ethanol by Pichia stipitis NCIM3498. Three different strategies for substrate hydrolysis using acid (dilute sulfuric acid) and alkali (dilute sodium hydroxide) and aqueous ammonia (AA) treatment followed by enzymatic hydrolysis were studied. A maximum of 631.5±3.25 mg/g sugars with 89.38% hydrolytic efficiency (HE) could be achieved after enzymatic hydrolysis of AA-pretreated S. spontaneum. All the substrate hydrolysates were evaluated for ethanol conversion in batches by P. stipitis. The microbial fermentation of released sugars into ethanol showed (g/g) 0.36±0.011, 0.384±0.022, 0.391±0.02, and 0.40±0.01 yield from detoxified acid hydrolysate and acid-, NaOH- and AA-pretreated substrate S. spontaneum enzymatic hydrolysates, respectively.