Biorefinery Gets Hot: Thermophilic Enzymes and Microorganisms for Second-Generation Bioethanol Production
Abstract
:1. Introduction
1.1. Need for Alternative Fuels
1.2. Bioethanol Biorefinery: State of the Art and Challenges
1.3. Advantages of Using Thermophiles in Biorefineries
2. Pretreatment of Lignocellulosic Biomasses
3. Enzymatic Hydrolysis
4. Thermophilic Fermentation
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Conflicts of Interest
References
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Enzyme [Reference] | Microorganism | Topt (°C) | pHopt | Thermal Stability (Activity %) | Substrate | Result |
---|---|---|---|---|---|---|
Endo-1,4-β-glucanaseAf-EGL7 [92] | Aspergillus fumigatus | 55 | 5 | 61% after 24 h at 60 °C | Corncob | The addition of 10 μg of Af-EGL7 to 0.009 FPU of Celluclast® 1.5 L increased the release of reducing sugars by 128% after 72 h (55 °C, pH 5) |
Rice straw | The addition of 10 μg of Af-EGL7 to 0.009 FPU of Celluclast® 1.5 L increased the release of reducing sugars by 80% after 72 h (55 °C, pH 5) | |||||
Endo-β-1,4-d-glucanase [93] | Trichoderma harzianum HZN11 | 60 | 5.5 | 66% after 3 h at 65 °C | Alkali pretreated sweet sorghum | 53 U/g mixed with 9 FP U/mL of commercial Trichoderma sps. increased the release of reducing sugars by 54% after 48 h (40 °C, pH 5.5) |
Sugarcane bagasse | 53 U/g mixed with 9 FP U/mL of commercial Trichoderma sps. increased the release of reducing sugars by 21% after 48 h (40 °C, pH 5.5) | |||||
Cellobiohydrolase CtCel7 [94] | Chaetomium thermophilum | 60 | 4 | 90% after 3 h at 60 °C | Pretreated wheat straw | The addition of CtCel7 to Sigma-Aldrich cellulase cocktail in 1:1 ratio increased the release of reducing sugars by 63% after 8 h (60 °C, pH 4). |
β-glucosidase [95] | Humicola grisea var. thermoidea | 50 | 6–7 | 50% after 7 and 14 min at 60 °C in absence or presence of 50 mM glucose | Sugarcane bagasse | The addition of 0.1 U of purified or crude β-glucosidase to 10 FPU T. reesei cellulases increased the saccharification by 50%. (50 °C, pH 5) |
β-glucosidase TN0602 [96] | Thermotoga naphthophila RUK-10 | 75 | 6 | 50% after 3 h at 90 °C [123] | Corn straw | A maximum increase in released glucose of 30.62% when 0.5 U/mL pf addition TN0602 were added to 0.75 U/mL of commercial cellulase from T. reesei (50 °C, pH 5) |
Endoxylanase (HXYN2) [97] | Humicola grisea var thermoidea, | 60 | 6.5 | 65% after 48 h at 50 °C | Steam-exploded corn straw | Addition of hemicellulases cocktail (600 U/g HXYN2, 11.5 U/g HXYLA and 0.32 U/g ABF3) to 5 FPU/g Accellerase 1500 enhanced the glucose yield by 14.6% in simultaneous reaction and by 50% in sequential reactions after 48 h, (50 °C, pH 5) |
β-xylosidase (HXYLA) [97] | 50 | 7 | 20% after 48 h at 50 °C | |||
α-l-arabinofuranosidase (AFB3) [97] | Penicillium pupurogenum | 50 | 5 | 25% after 48 h at 50 °C | ||
XynZ-C and Xyn11A [98] | Clostridium thermocellum Thermobifida fusca | 75 | 7 | 96% after 18h at 75 °C in 10% glycerol [124] | Pretreated bagasse | 2-fold increase in the concentration of reducing sugars, when both xylanases, were added to T. reesei cellulase in 50:50 ratio (50 °C, pH 6) |
Xylanase (TcXyn10A) [99] | Thermobacillus composti | 65 | 6–8 | 40% after 8 h at 65 °C | Sugarcane bagasse | Improvement of hydrolysis Accellerase® 1500 cocktail hydrolysis by 15.35 % increase in xylose release and 4.38% glucose release after 24 h in 1:1 ratio (50 °C pH 5) |
Endo-β-mannanase MtMan26A [100] | Myceliophthora thermophila | 60 | 6 | 50% after 14.4 h at 60 °C | Pretreated beechwood sawdust | Release of total reducing sugars and glucose improved by 13 and 12%, as a supplement to Celluclast® 1.5 L and Novozyme® 188 in 1:1 ratio (50 °C pH 5) |
LPMO PMO9D_SCYTH [104] | Scytalidium thermophilum | 60 | 7 | 50% after 60.58 h at 60 °C | Bagasse | 18.9 and 17.5% yield increase from acid or alkali-treated bagasse respectively (Cellic CTec2, 9:1 ratio, 50 °C pH 5) |
Rice straw | 28.7 and 22.1% yield increase from acid or alkali-treated rice straw respectively (Cellic CTec2, 9:1 ratio, 50 °C pH 5) | |||||
LPMO PMO9D_MALCI [104] | Malbranchea cinnamomea | 50 | 9 | 50 after 144 h at 50 °C | Bagasse | 21.3 and 23.6% yield increase from acid or alkali-treated bagasse respectively (Cellic CTec2, 9:1 ratio, 50 °C pH 5) |
Rice straw | 28.8 and 13.6% yield increase from acid or alkali respectively (Cellic CTec2, 9:1 ratio, 50 °C pH 5) |
Strain [Reference] | Feedstock | Temperature (°C) | Total Fermentation Time (a) | Process Scheme | Maximum Ethanol Titer (b) | Maximum Ethanol Yield (c) |
---|---|---|---|---|---|---|
K. marxianus MTCC 1389 [168] | Prosopis juliflora woody stems | 41 | 72 h (72 h) | SSF | 21.45 g/L | 0.67 g/g |
K. marxianus ATCC 36,907 [169] | Cashew apple bagasse | 40 | 72 h (~28 h) | SSF | 68 g/L | 80.70% |
K. marxianus S1.17 [170] | Sugarcane leaves | 40 | 132 h (~48 h) | SSF | 5.59 g/L | 0.10 g/g dry weight |
K. marxianus K213 [133] | Water hyacinth | 42 | 24 h (24 h) | SSF | 7.34 g/L | 0.16 g/g biomass |
K. marxianus KCTC7001 [171] | Empty palm fruit bunches | 42 | ~28 h | SSF | 7.80% | |
K. marxianus ATCC 36,907 [172] | Carnauba straw residue | 45 | 48 h (12 h) | SSF | 7.52 g/L | 75.29% |
K. marxianus ATCC 36,907 [132] | Sugarcane bagasse | 43 | 24 h | SSF | 4.18 g/100 g biomass | |
K. marxianus TY16 [173] | Bamboo | 42.5 | 108 h (108 h) | SSF | 26.04 g/L | |
C. thermocellum DSM 1313 [136] | Rice husk | 60 | 120 h | CBP | 1 g/L | |
C. thermocellum [136] | Sugarcane bagasse | 60 | 120 h | CBP | 1.21 g/L | |
C. thermocellum ATCC 27,405 and T. thermosaccharolyticum DSM 571 [150] | Corn straw | 55 | 168 h (120 h) | CBP | 0.45 g/L | 11.20% |
C. thermocellum ATCC 27,405 and T. thermosaccharolyticum DSM 571 [151] | Salix | 55 | 168 h | CBP | 0.2 g/L | 11.10% |
C. thermocellum CT2 and Clostridium thermosaccharolyticum HG8 [174] | Banana Agro-waste | 60 | 120 h | CBP | 0.41 g/g substrate | |
C. thermocellum [152] | Sugarcane bagasse | 55 | 168 h | CBP | 10.60 mM | |
Clostridium sp. DBT-IOC-C19 and Thermoanaerobacter sp. DBT-IOC-X2 [153] | Rice straw slurry | 60 | 144 h (144 h) | CBP | 142 mM | 48% |
C. thermocellum DSM 1237 [154] | Sugarcane bagasse | 60 | 60 h (~28 h) | CBP | 0.86 g/L | 83.30% |
Engineered C. bescii [156] | Switchgrass | 75 | 50 h (15 h) | CBP | ~2 mM | |
C. bescii MACB 1058 [158] | Poplar (transgenic) | 65 | 168 h | CBP | 18.3 mM | |
G. thermoglucosidasius and T. ethanolicus [162] | Food waste | 60 | 120 h (120 h) | CBP | 18.4 g/L | 0.24 g/g sugar |
Strain [Reference] | Protein Function (Gene) | Source Organism | Effect |
---|---|---|---|
C. thermocellum DSM1313 [142] | Xylose isomerase (xylA), xylulokinase (xylB). | T. ethanolicus | Enabled xylose co-fermentation. |
C. thermocellum DSM1313 [143] | Bifunctional alcohol dehydrogenases (adhE), NADH-dependent reduced ferredoxin:NADP+ oxidoreductase complex (nfnA/B), and NADPH- dependent alcohol dehydrogenase (adhA) | T. saccharolyticum | Enhanced ethanol production. |
C. thermocellum DSM1313 Δhpt ΔhydG [163] | Lactate dehydrogenase (ldh), pyruvate-formate lyase (pfl), and phosphotransacetylase and acetate kinase (pta-ack) | Self, knockout | Enhanced ethanol production. |
C. thermocellum [164] | Gglyceraldehyde 3-phosphate dehydrogenase (gapdh) | T. saccharolyticum | Enhanced ethanol tolerance. |
C. thermocellum [165] | Spermidine synthase (speE) | Self, overexpression | Enhanced acetate and HMF tolerance. |
C. thermocellum [175] | Ion-translocating reduced ferredoxin: NAD+ oxidoreductase (rnf) and hydrogenase maturation gene (hydG) | Self, overexpression, and knockout, respectively | Enhanced ethanol production. |
T. thermosaccharolyticum [138] | A-d-xylosidase (α-Xylp_1211), α-l-galactosidases (α-l-Galp_687 and 697), β-d-xylosidase (β-Xylp_1710), and α-l-arabinofuranosidases (α-Araf_996 and 1120) | Herbinix spp. | Conferred ability to deconstruct GAX component of corn fiber |
T. saccharolyticum, T. thermosaccharolyticum, T. xylanolyticum, T. mathranii, and C. thermocellum [176] | Hydrogenase (hfsB) | Self, knockout | Enhanced ethanol production. |
Thermoanaerobacterium aotearoense SCUT27/Δldh [177] | Pyruvate formate lyase-activating protein A (pflA) | Self, knockout | Enhanced ethanol tolerance and production |
C. bescii Δldh [156] | Bi-functional acetyl-CoA thioesterase/ alcohol dehydrogenase (adhB) and bi-functional acetaldehyde/alcohol dehydrogenase (adhE) | Thermoanaerobacter pseudethanolicus 39E | Enabled ethanol fermentation at high temperatures. |
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Zuliani, L.; Serpico, A.; De Simone, M.; Frison, N.; Fusco, S. Biorefinery Gets Hot: Thermophilic Enzymes and Microorganisms for Second-Generation Bioethanol Production. Processes 2021, 9, 1583. https://doi.org/10.3390/pr9091583
Zuliani L, Serpico A, De Simone M, Frison N, Fusco S. Biorefinery Gets Hot: Thermophilic Enzymes and Microorganisms for Second-Generation Bioethanol Production. Processes. 2021; 9(9):1583. https://doi.org/10.3390/pr9091583
Chicago/Turabian StyleZuliani, Luca, Annabel Serpico, Mario De Simone, Nicola Frison, and Salvatore Fusco. 2021. "Biorefinery Gets Hot: Thermophilic Enzymes and Microorganisms for Second-Generation Bioethanol Production" Processes 9, no. 9: 1583. https://doi.org/10.3390/pr9091583