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  • 1.
    Nair, Ramkumar B
    University of Borås, Faculty of Textiles, Engineering and Business.
    Integration of first and second generation bioethanol processes using edible filamentous fungus Neurospora intermedia2017Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    Establishing a commercial, lignocellulose-based, second-generation ethanol process has received several decades of attention by both researchers and industry. However, a fully economically viable process still remains a long-term goal. The main bottleneck to this achievement is the recalcitrance of lignocellulosic feedstocks, although there are several other factors, such as the huge investment required for second-generation ethanol facilities. An intelligent alternative solution discussed in this thesis is an integrated approach using firstgeneration ethanol plants for second-generation processes. Wheat is the major feedstock for first-generation ethanol in Europe; therefore, wheat-based lignocellulose waste, such as wheat straw, bran, and whole stillage fiber (a waste stream from first-generation wheat-based ethanol plants) was the primary focus of the integration model in this thesis. Since the major share of first-generation ethanol plant economics focuses on the animal feed DDGS (Distillers’ dried gains with solubles), the integration of lignocellulose should be designed in order to maintain DDGS quality. An ethanol-producing edible filamentous fungus, Neurospora intermedia, a potential protein source in DDGS, was considered for use as the fermenting microbe. The morphological and physiological aspects of this fungus were studied in the thesis, leading to the first report of fungal pellet development.

    An alternative approach of using dilute phosphoric acid to pretreat lignocellulose, as it does not negatively affect fungal growth or DDGS quality, was demonstrated in both the laboratory and on a 1m3 pilot scale. Furthermore, the process of hydrolysis of pretreated lignocelluloses and subsequent N. intermedia fermentation on lignocellulose hydrolysate was also optimized in the laboratory and scaled up to 1 m3 using an in-house pilot-scale airlift bioreactor. Fungal fermentation on acid-pretreated and enzyme-hydrolyzed wheat bran, straw and whole stillage fiber resulted in a final ethanol yield of 95%, 94% and 91% of the theoretical maximum based on the glucan content of the substrate, respectively. Integrating the first- and second-generation processes using thin stillage (a waste stream from first-generation wheat-based ethanol plants) enhanced the fungal growth on straw hydrolysate, avoiding the need for supplementing with extra nutrients.

    Based on the results obtained from this thesis work, a new model for integrated first- and second-generation ethanol using edible filamentous fungi processes that also adds value to animal feed (DDGS) was developed.

  • 2.
    Nair, Ramkumar B
    University of Borås, Faculty of Textiles, Engineering and Business. Biotechnology.
    Ferreira, Jorge
    University of Borås, Faculty of Textiles, Engineering and Business. Biotechnology.
    Taherzadeh, Mohammad
    University of Borås, Faculty of Textiles, Engineering and Business. Biotechnology.
    Lennartsson, Patrik
    University of Borås, Faculty of Textiles, Engineering and Business. Biotechnology.
    Mild-temperature dilute acid pretreatment for integration of first and second generation ethanol processes2017In: Bioresource Technology, ISSN 0960-8524, E-ISSN 1873-2976, Vol. 245, 145-151 p.Article in journal (Refereed)
    Abstract [en]

    The use of hot-water (100 °C) from the 1st generation ethanol plants for mild-temperature lignocellulose pretreatment can possibly cut down the operational (energy) cost of 2nd generation ethanol process, in an integrated model. Dilute-sulfuric and -phosphoric acid pretreatment at 100 °C was carried out for wheat bran and whole-stillage fibers. Pretreatment time and acid type influenced the release of sugars from wheat bran, while acid-concentration was found significant for whole-stillage fibers. Pretreatment led up-to 300% improvement in the glucose yield compared to only-enzymatically treated substrates. The pretreated substrates were 191–344% and 115–300% richer in lignin and glucan, respectively. Fermentation using Neurospora intermedia, showed 81% and 91% ethanol yields from wheat bran and stillage-fibers, respectively. Sawdust proved to be a highly recalcitrant substrate for mild-temperature pretreatment with only 22% glucose yield. Both wheat bran and whole-stillage are potential substrates for pretreatment using waste heat from the 1st generation process for 2nd generation ethanol.

  • 3.
    Nair, Ramkumar B
    et al.
    University of Borås, School of Engineering.
    Brandberg, Thomas
    Lennartsson, Patrik R
    University of Borås, School of Engineering.
    Taherzadeh, Mohammad J
    University of Borås, School of Engineering.
    Dilute phosphoric acid pretreatment of wheat straw for ethanol production using edible fungi2014Conference paper (Refereed)
  • 4.
    Nair, Ramkumar B
    et al.
    University of Borås, Faculty of Textiles, Engineering and Business.
    Kabir, Maryam M.
    Lennartsson, Patrik R.
    University of Borås, Faculty of Textiles, Engineering and Business.
    Taherzadeh, Mohammad J
    University of Borås, Faculty of Textiles, Engineering and Business.
    Sárvári Horváth, Ilona
    University of Borås, Faculty of Textiles, Engineering and Business.
    Integrated Process for Ethanol, Biogas, and Edible Filamentous Fungi-Based Animal Feed Production from Dilute Phosphoric Acid-Pretreated Wheat Straw2017In: Applied Biochemistry and Biotechnology, ISSN 0273-2289, E-ISSN 1559-0291, 1-15 p.Article in journal (Refereed)
    Abstract [en]

    Integration of wheat straw for a biorefinery-based energy generation process by producing ethanol and biogas together with the production of high-protein fungal biomass (suitable for feed application) was the main focus of the present study. An edible ascomycete fungal strain Neurospora intermedia was used for the ethanol fermentation and subsequent biomass production from dilute phosphoric acid (0.7 to 1.2% w/v) pretreated wheat straw. At optimum pretreatment conditions, an ethanol yield of 84 to 90% of the theoretical maximum, based on glucan content of substrate straw, was observed from fungal fermentation post the enzymatic hydrolysis process. The biogas production from the pretreated straw slurry showed an improved methane yield potential up to 162% increase, as compared to that of the untreated straw. Additional biogas production, using the syrup, a waste stream obtained post the ethanol fermentation, resulted in a combined total energy output of 15.8 MJ/kg wheat straw. Moreover, using thin stillage (a waste stream from the first-generation wheat-based ethanol process) as a co-substrate to the biogas process resulted in an additional increase by about 14 to 27% in the total energy output as compared to using only wheat straw-based substrates. .[on SciFinder (R)]

  • 5.
    Nair, Ramkumar B.
    et al.
    University of Borås, Faculty of Textiles, Engineering and Business.
    Lundin, Magnus
    University of Borås, Faculty of Textiles, Engineering and Business.
    Lennartsson, Patrik R.
    University of Borås, Faculty of Textiles, Engineering and Business.
    Taherzadeh, Mohammad J
    University of Borås, Faculty of Textiles, Engineering and Business.
    Optimizing dilute phosphoric acid pretreatment of wheat straw in the laboratory and in a demonstration plant for ethanol and edible fungal biomass production using Neurospora intermedia.2016In: Journal of chemical technology and biotechnology (1986), ISSN 0268-2575, E-ISSN 1097-4660Article in journal (Refereed)
    Abstract [en]

    BACKGROUND : A method is described that uses dil. phosphoric acid for wheat straw pretreatment and subsequent ethanol and fungal biomass prodn. with the edible fungus Neurospora intermedia. Dil. phosphoric acid pretreatment of wheat straw was optimized at a lab. scale, and the results were validated in a biorefinery demonstration plant for the first time. The various conditions for the dil. acid pretreatment include such factors as phosphoric acid concns. (0.5-3.0% w/v), temp. (150-210 °C), and reaction time (5-20 min). RESULTS : The optimal pretreatment conditions were detd. as an acid concn. of 1.75% (w/v) at a temp. of 190 °C for 15 min, based on the max. enzymic digestibility with the min. inhibitor release. The efficiency of enzymic polysaccharide hydrolysis was 36% for untreated straw and 86% for straw pretreated with dil. phosphoric acid. Scale up of the pretreatment at a biorefinery demonstration plant improved the process, with the subsequent efficiency of polysaccharide hydrolysis being 95% of the theor. max. Ethanol fermn. of enzymically hydrolyzed wheat straw using N. intermedia showed an improvement in the ethanol yield from 29% (with untreated straw) to 94% (with dil. phosphoric acid pretreated straw) of the theor. max. CONCLUSION : This study opens up an alternative strategy for the efficient use of wheat straw for the prodn. of ethanol and edible fungal biomass in existing wheat-to-ethanol plants.

  • 6.
    Nair, Ramkumar B
    et al.
    University of Borås, Faculty of Textiles, Engineering and Business. Swedish Centre for Resource Recovery.
    Ravula, Vamsikrishna
    University of Borås, Faculty of Textiles, Engineering and Business. Swedish Centre for Resource Recovery.
    Lennartsson, Patrik
    University of Borås, Faculty of Textiles, Engineering and Business. Swedish Centre for Resource Recovery.
    Taherzadeh, Mohammad
    University of Borås, Faculty of Textiles, Engineering and Business. Swedish Centre for Resource Recovery.
    Neurospora intermedia pellets for enhanced ethanol and fungal biomass production from wheat straw2017In: Proceedings of 39th Symposium on Biotechnology for Fuels and Chemicals, 2017Conference paper (Refereed)
    Abstract [en]

    Recent studies at our research group have described an ‘integrated-biorefinery’ model for the existing 1st generation wheat-based ethanol facilities, by using edible filamentous fungus, Neurospora intermedia. The process focuses on the production of 2nd generation ethanol together with fungal biomass (for animal or aquaculture feed applications) from wheat straw. A final ethanol yield of 94% (theoretical maximum based on substrate glucan content) was obtained with N. intermedia fermentation in dilute phosphoric acid pretreated (0.7%w/v acid, 7min at 201±4°C) and enzymatically hydrolyzed (10FPU cellulase/g substrate) straw. Fungal cultivation in liquid straw hydrolysate resulted in a maximum of 3.71±0.11g/L dry fungal biomass. Considering the industrial significance of the fungal process, attempts were made to manipulate N. intermedia to grow as pellet forms in the straw hydrolysate, for the first time. Of the various culture conditions screened, stable pellet morphology was obtained at pH 3.0 to 5.5, resulting in uniform pellets with size ranging from 2.5 to 4.25mm. Fermentation using N. intermedia pellets in the liquid straw hydrolysate, resulted in about 31% increase in the ethanol yield, with an improved glucose assimilation by the pellets (82% reduction) as opposed to filamentous forms (51% reduction), at similar culture conditions. The growth of fungal pellets in presence of inhibitors (at different concentrations of acetic acid and furfural) resulted in about 11% to 45% increase in ethanol production as compared to filamentous forms, at similar growth conditions in the liquid straw hydrolysate. Detailed results on N. intermedia pelletization in liquid straw hydrolysate will be discussed in this presentation.

  • 7.
    Nair, Ramkumar B
    et al.
    University of Borås, Faculty of Textiles, Engineering and Business.
    Taherzadeh, Mohammad J
    University of Borås, Faculty of Textiles, Engineering and Business.
    Process for edible filamentous fungal cultivation and its integration in conventional sugar to ethanol production.2017Patent (Other (popular science, discussion, etc.))
    Abstract [en]

    The present invention relates to a process for edible fungal cultivation by employing vinasse or spent wash of first generation ethanol prodn. as a substrate. More particularly, the present invention relates to a process for edible fungal cultivation in the form of pellets. [on SciFinder(R)]

  • 8.
    Nair, Ramkumar
    et al.
    University of Borås, Faculty of Textiles, Engineering and Business. Biotechnology.
    Eh-Hser Nay, Theimya
    Biotechnology.
    Lennartsson, Patrik
    University of Borås, Faculty of Textiles, Engineering and Business. Biotechnology.
    Taherzadeh, Mohammad
    University of Borås, Faculty of Textiles, Engineering and Business. Biotechnology.
    Waste Bread Valorization Using Edible Filamentous Fungi2017Conference paper (Refereed)
    Abstract [en]

    The present study is the first of its kind to use industrial waste bread for ethanol and food-grade filamentous fungal biomass production, with an ‘integrated-biorefinery’ approach for the existing wheat-based ethanol facilities. Four different food-grade fungi such as Neurospora intermedia, Aspergillus oryzae, belonging to ascomycetes and Mucor indicus, Rhizopus oryzae, belonging to zygomycetes, were screened. Initial screening for fungal cultures (without external enzyme saccharification) showed an ethanol yield maximum of 47.8 ±1.1 to 67.3 ±2.1, and 38.7 ±1.1 to 67.7±1.8 mg per g dry substrate loading from whole-grain bread and white-bread respectively, post the enzymatic liquefaction. Scale-up of the N. intermedia fermentation achieved using bench scale airlift reactor showed an ethanol yield maximum of 91.6 ±2.1 and 87.5 ±1.9 mg per g dry substrate loading for whole-grain bread and white-bread respectively.

  • 9.
    Nair, Ramkumar
    et al.
    University of Borås, Faculty of Textiles, Engineering and Business. Swedish Centre for Resource Recovery.
    Ferreira, Jorge
    University of Borås, Faculty of Textiles, Engineering and Business. Swedish Centre for Resource Recovery.
    Lennartsson, Patrik R.
    University of Borås, Faculty of Textiles, Engineering and Business. Swedish Centre for Resource Recovery.
    Taherzadeh, Mohammad J
    University of Borås, Faculty of Textiles, Engineering and Business.
    Evaluating dilute phosphoric acid pretreated wheat bran for ethanol and edible fungal biomass production: A pilot study2015Conference paper (Refereed)
    Abstract [en]

    Purpose of the work:

    The present study reports the application of using dilute phosphoric acid for pretreatment of wheat branbiomass and its subsequent use as an ethanol fermentation substrate for edible fungi, Neurospora intermedia inbench scale (4.5L) and pilot airlift (26L) reactors.

    Approach:

    The optimized pretreatment conditions to release maximum sugar and minimum inhibitor concentrations weredetermined using a statistically modelled experimental setup. The dilute acid pretreatment was scaled up withthe validation of lab results in a biorefinery demo plant (BDP) at Svensk Etanolkemi AB – SEKAB (Örnsköldsvik,Sweden). The structural and physiological changes in the bran biomass during the pretreatment process werealso studied. Bench scale and pilot scale enzymatic hydrolysis of the pretreated biomass and subsequent ethanolfermentation using edible ascomycetes fungi, N. intermedia were studied using airlift reactors.

    Scientific innovation and relevance:

    With the use of dilute phosphoric acid, the study put forth an alternative strategy to the current use of relativelystronger acids for lignocellulosic biomass pretreatment. The potential use of acid pretreated lignocellulosicbiomass (wheat bran) as an ethanol feedstock, together with the edible fungal cultivation is a first of its kindapproach towards a wider ‘biorefinery’ concept. Improving the co-product (DDGS) quality and ethanol yield inthe existing wheat based ethanol facilities, whereby enhancing the overall ethanol production economics formsthe ultimate aim of the study.

    Results:

    Wheat bran was subjected to dilute acid pretreatment at varying acid concentrations (0.5–3.0% w/v),temperature (150–210◦C), and reaction time (5–20 min). The interaction of multiple factors showed theoptimum pretreatment conditions at acid concentration of 1.75% (w/v), at 190◦C for 10 min. A maximum totalpolysaccharide yield of 0.27 ± 0.01 g/g dry biomass loading, corresponding to 66% of the theoretical maximumwas observed. The effect of the dilute acid pretreatment on the functional groups of the wheat bran cellulosewas determined with 78% reduction in the cellulose crystallinity index. Enzymatic hydrolysis of pretreated slurryfrom the demo plant showed 85% total theoretical yield of polysaccharides. Compared to the untreated branbiomass, an increase of 51% was observed in the ethanol yield following pretreatment, with a total ethanol yieldof 95% theoretical maximum. Similar results were also observed on scaling up the reaction volume in a benchscale (4.5L) and pilot scale (26L) airlift reactors, which will also be presented in detail.

    Conclusions:

    The present work brings out the potential of using phosphoric acid as an efficient pretreatment agent addressingthe long existing problems associated with the sulfur contamination of the feed. Additional advantages relatedto the nutritional enrichment of DDGS (at the existing wheat based ethanol facility), with the use of edibleascomycetes fungi were also achieved. However, techno-economical analyses are required to determine theactual industrial feasibility of the process.

  • 10.
    Nair, Ramkumar
    et al.
    University of Borås, Faculty of Textiles, Engineering and Business.
    Lennartsson, Patrik R.
    University of Borås, Faculty of Textiles, Engineering and Business.
    Taherzadeh, Mohammad J
    University of Borås, Faculty of Textiles, Engineering and Business.
    Bioethanol production from agricultural and municipal waste2016In: Current Developments in Biotechnology and Bioengineering: Solid Waste Management / [ed] Wong, J.; Tyagi, R.D.; Pandey, A., USA: Elsevier, 2016, 1, 157-190 p.Chapter in book (Refereed)
    Abstract [en]

    Bioethanol, one of the most promising technological advancements of the century, has been widely acclaimed for being produced from diversified origins. Production of bioethanol from food grains (as in Brazil or the United States) is however frequently criticized in the food vs. fuel debate. Several research studies across the globe, investigating the potential use of various renewable resources (such as waste biomass), have resulted in the emergence of second and/or third generation bioethanol processes. This chapter attempts to consolidate various aspects of bioethanol production from solid waste biomass. Waste biomass of lignocellulosic and starch-based origin, such as municipal solid waste, industrial waste (waste paper or coffee residues), livestock manure, and agricultural waste (wood biomass and agricultural crop residues), were reviewed for their potential to produce ethanol. This chapter describes the feedstock prospects, process technologies, and the current research and industrial developments.

  • 11.
    Nair, Ramkumar
    et al.
    University of Borås, Faculty of Textiles, Engineering and Business.
    Lennartsson, Patrik
    University of Borås, Faculty of Textiles, Engineering and Business.
    Taherzadeh, Mohammad
    University of Borås, Faculty of Textiles, Engineering and Business.
    Mycelial pellet formation by edible ascomycete filamentous fungi, Neurospora intermedia2016In: AMB Express, ISSN 2191-0855, E-ISSN 2191-0855, Vol. 6, no 31, 10.1186/s13568-016-0203-2Article in journal (Refereed)
    Abstract [en]

    Pellet formation of filamentous fungi in submerged culture is an imperative topic of fermentation research. In this study, we report for the first time the growth of filamentous ascomycete fungus,Neurospora intermedia in its mycelial pellet form. In submerged culture, the growth morphology of the fungus was successfully manipulated into growing as pellets by modifying various cultivation conditions. Factors such as pH (2.0–10.0), agitation rate (100–150 rpm), carbon source (glucose, arabinose, sucrose, and galactose), the presence of additive agents (glycerol and calcium chloride) and trace metals were investigated for their effect on the pellet formation. Of the various factors screened, uniform pellets were formed only at pH range 3.0–4.0, signifying it as the most influential factor for N. intermedia pellet formation. The average pellet size ranged from 2.38 ± 0.12 to 2.86 ± 0.38 mm. The pellet formation remained unaffected by the inoculum type used and its size showed an inverse correlation with the agitation rate of the culture. Efficient glucose utilization was observed with fungal pellets, as opposed to the freely suspended mycelium, proving its viability for fast- fermentation processes. Scale up of the pelletization process was also carried out in bench-scale airlift and bubble column reactors (4.5 L).

  • 12.
    Nair, Ramkumar
    et al.
    University of Borås, Faculty of Textiles, Engineering and Business. Swedish Centre for Resource Recovery.
    Lundin, Magnus
    University of Borås, Faculty of Textiles, Engineering and Business. Swedish Centre for Resource Recovery.
    Brandberg, Tomas
    Lennartsson, Patrik
    University of Borås, Faculty of Textiles, Engineering and Business. Swedish Centre for Resource Recovery.
    Taherzadeh, Mohammad J
    University of Borås.
    Dilute phosphoric acid pretreatment of wheat bran for enzymatic hydrolysis and subsequent ethanol production by edible fungi Neurospora intermedia.2015In: Industrial crops and products (Print), ISSN 0926-6690, E-ISSN 1872-633X, Vol. 69, 314-323 p.Article in journal (Refereed)
    Abstract [en]

    The use of an underutilized and abundant lignocellulosic feedstock residue, wheat bran, was studied for ethanol prodn. using dil. phosphoric acid pretreatment followed by fermn. using edible fungi Neurospora intermedia. Wheat bran was subjected to dil. acid pretreatment at varying acid concns. (0.5-3.0% w/v), temp. (150-210 °C), and reaction time (5-20 min). The interaction of multiple factors showed the optimum pretreatment conditions at acid concn. of 1.75% (w/v), at 190 °C for 10 min. The max. total polysaccharide yield of 0.27 ± 0.01 g/g dry biomass loading, corresponding to 66% of the theor. max. was obsd. Subsequent fermn. with N. intermedia showed 85% of the theor. max. ethanol yield from the untreated bran glucose. The effect of the dil. acid pretreatment on the functional groups of the wheat bran cellulose was detd. with 78% redn. in the cellulose crystallinity index. The validation of the dil. phosphoric acid pretreatment in a demo plant is also reported for the first time. Enzymic hydrolysis of pretreated slurry from the demo plant showed 85% total theor. yield of polysaccharides. Compared to the untreated bran biomass, an increase of 51% was obsd. in the ethanol yield following pretreatment, with a total ethanol yield of 95% theor. max. Higher yield of ethanol is also attributed to the xylose fermenting capability of the fungi. [on SciFinder(R)]

  • 13.
    Nair, Ramkumar
    et al.
    University of Borås, Faculty of Textiles, Engineering and Business.
    Taherzadeh, Mohammad J
    University of Borås, Faculty of Textiles, Engineering and Business.
    Valorization of sugar-to-ethanol process waste vinasse: A novel biorefinery approach using edible ascomycetes filamentous fungi2016In: Bioresource Technology, ISSN 0960-8524, E-ISSN 1873-2976, Vol. 221, 469-476 p.Article in journal (Refereed)
    Abstract [en]

    The aim of the present work was to study the integration of edible ascomycetes filamentous fungi into the existing sugar- or molasses-to-ethanol processes, to grow on vinasse or stillage and produce ethanol and protein-rich fungal biomass. Two fungal strains, Neurospora intermedia and Aspergillus oryzae were examined in shake flasks and airlift-bioreactors, resulting in reduction of vinasse COD by 34% and viscosity by 21%. Utilization of glycerol and sugars were observed, yielding 202.4 or 222.8g dry fungal biomass of N. intermedia or A. oryzae respectively, per liter of vinasse. Integration of the current process at an existing ethanol facility producing about 100,000m(3) of ethanol per year could produce around 200,000-250,000tons of dry fungal biomass (40-45% protein) together with about 8800-12,600m(3) extra ethanol (8.8-12.6% of production-rate improvement).[on SciFinder (R)]

  • 14. Osadolor, Osagie A.
    et al.
    Nair, Ramkumar B
    University of Borås, Faculty of Textiles, Engineering and Business.
    Lennartsson, Patrik R.
    University of Borås, Faculty of Textiles, Engineering and Business.
    Taherzadeh, Mohammad J
    University of Borås, Faculty of Textiles, Engineering and Business.
    Empirical and experimental determination of the kinetics of pellet growth in filamentous fungi: A case study using Neurospora intermedia.2017In: Biochemical engineering journal, ISSN 1369-703X, E-ISSN 1873-295X, Vol. 124, 115-121 p.Article in journal (Refereed)
    Abstract [en]

    Pellet morphol. formation by filamentous fungi has gained a lot of attention because of its multiple benefits such as the ease of sepn. and smaller bioreactor vol. requirement. Most reported kinetics studies on fungal pellet growth are centered on aeration, despite the exptl. results pointing to the importance of other factors such as pH, substrates and product concn. etc., influencing the pellet formation. Hence a kinetic study on the effect of multiple factors such as aeration, substrate and product concn. and pH was done in this paper using Neurospora intermedia as a model organism, whose ability to form mycelial pellets was recently reported. The max. growth rate of the pellets under uninhibited conditions at its optimal growth pH was 0.318 h-1. The pellets were found to be inhibited by high product (ethanol) concn. with no growth occurring at 70 g L-1 and above. High substrate concn. favored the formation of loose fur-like fluffy pellets. The specific oxygen uptake rate of the pellets was between 0.27-0.9 mmol-O2 g-biomass-1h-1 depending on the pellet av. diam. The results from this kinetic study can be used for bioreactor design, operations and optimization of fermn. processes utilizing N. intermedia. [on SciFinder(R)]

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