Abstract
Microalgae species including Chlamydomonas mexicana, Micractinium reisseri, Scenedesmus obliquus and Tribonema aequale were cultivated in batch cultures, and their biochemical composition was determined. C. mexicana showed the highest carbohydrate content of 52.6 % and was selected for further study. Sonication pretreatment under optimum conditions (at 40 kHz, 2.2 Kw, 50 °C for 15 min) released 74 ± 2.7 mg g−1 of total reducing sugars (TRS) of dry cell weight, while the combined sonication and enzymatic hydrolysis treatment enhanced the TRS yield by fourfold (280.5 ± 4.9 mg g−1). The optimal ratio of enzyme [E]:substrate [S] for maximum TRS yield was [1]:[5] at 50 °C and pH 5. Combined sonication and hydrolysis treatment released 7.3 % (27.1 ± 0.9 mg g−1) soluble protein of dry cell weight, and further fermentation of the dissolved carbohydrate fraction enhanced the soluble protein content up to 56 % (228.4 mg g−1) of total protein content. Scanning and transmission electron microscopic analyses indicated that microalgae cells were significantly disrupted by the combined sonication and enzyme hydrolysis treatment. This study indicates that pretreatment and subsequent fermentation of the microalgal biomass enhance the recovery of carbohydrates and proteins which can be used as feedstocks for generation of biofuels.
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Hernandez D, Riano B, Coca M, Garcia-Gonzalez MC (2015) Saccharification of carbohydrates in microalgal biomass by physical, chemical and enzymatic pre-treatments as a previous step for bioethanol production. Chem Eng J 262:939–945
Scholz MJ, Riley MR, Cuello JL (2013) Acid hydrolysis and fermentation of microalgal starches to ethanol by the yeast Saccharomyces cerevisiae. Biomass Bioenerg 48:59–65
Kalyani D, Tiwari MK, Li J, Kim SC, Kalia VC, Kang YC (2015) A Highly Efficient Recombinant Laccase from the Yeast Yarrowia lipolytica and Its Application in the Hydrolysis of Biomass. PLoS One 10(3):e0120156
Chisti Y (2008) Biodiesel from microalgae beats bioethanol. Trends Biotechnol 26:126–131
Hu Q, Kurano N, Kawachi M, Iwasaki I, Miyachi S (1998) Ultrahigh-cell-density culture of a marine green alga Chlorococcum littorale in a flat-plate photobioreactor. Appl Microbiol Biotechnol 49:655–662
Halim R, Harun R, Danquah MK, Webley PA (2012) Microalgal cell disruption for biofuel development. Appl Energy 91:116–121
Taherzadeh MJ, Karimi K (2007) Enzymatic-based hydrolysis processes for ethanol from lignocellulosic materials a review. BioResources 2(4):707–738
John RP, Anisha GS, Nampoothiri KM, Pandey A (2011) Micro and macroalgal biomass: a renewable source for bioethanol. Bioresour Technol 102:186–193
Ho SH, Huang SW, Chen CY, Hasunuma T, Kondo A, Chang JS (2013) Bioethanol production, using carbohydrate-rich microalgae biomass as feedstock. Bioresour Technol 135:191–198
Lee JY, Yoo C, Jun SY, Ahn CY, Oh HM (2010) Comparison of several methods for effective lipid extraction from microalgae. Bioresour Technol 101:75–77
Kashaninejad M, Tabil LG (2011) Effect of microwave-chemical pre-treatment on compression characteristics of biomass grinds. Biosyst Eng 108:36–45
Wang XQ, Zhang XW (2012) Optimal extraction and hydrolysis of Chlorella pyrenoidosa proteins. Bioresour Technol 126:307–313
Hernandez D, Solana M, Riano B, Garcia-Gonzalez MC, Bertucco A (2014) Biofuels from microalgae: lipid extraction and methane production from the residual biomass in a biorefinery approach. Bioresour Technol 170:370–378
Sulaiman AZ, Ajit A, Chisti Y (2013) Ultrasound mediated enzymatic hydrolysis of cellulose and carboxymethyl cellulose. Biotechnol Prog 29:1448–1457
Jeon B-H, Choi JA, Kim HC, Hwang J-H, Abou-Shanab RAI, Dempsey BA, Regan JM, Kim JR (2013) Ultrasonic disintegration of microalgal biomass and consequent improvement of bioaccessibility/bioavailability in microbial fermentation. Biotechnol Biofuels 6(37):1–9. doi:10.1186/1754-6834-6-37
Choi JA, Hwang JH, Dempsey BA, Abou-Shanab RAI, Min B, Song H, Lee DS, Kim JR, Cho Y, Hong S, Jeon B-H (2011) Enhancement of fermentative bioenergy (ethanol/hydrogen) production using ultrasonication of Scenedesmus obliquus YSW15 cultivated in swine wastewater effluent. Energy Environ Sci 4:3513–3520
Gonzalez-Fernandez C, Sialve B, Bernet N, Steyer JP (2012) Comparison of ultrasound and thermal pretreatment of Scenedesmus biomass on methane production. Bioresour Technol 110:610–616
Bhatia L, Johri S, Ahmad R (2012) An economic and ecological perspective of ethanol production from renewable agro waste: a review. AMB Express 2:65–84
Kalyani D, Lee KM, Kim TS, Li J, Dhiman SS, Kang YC, Lee JK (2013) Microbial consortia for saccharification of woody biomass and ethanol fermentation. Fuel 107:815–822
Tan IS, Lee KT (2014) Enzymatic hydrolysis and fermentation of seaweed solid wastes for bioethanol production: an optimization study. Energy 78:53–62
Kuhad RC, Gupta R, Khasa YP, Singh A, Zhang YHP (2011) Bioethanol production from pentose sugars: current status and future prospects. Renew Sust Energy Rev 15:4950–4962
Fu CC, Hung TC, Chen JY, Su CH, Wu WT (2010) Hydrolysis of microalgae cell walls for production of reducing sugar and lipid extraction. Bioresour Technol 101:8750–8754
Singh S, Khanna S, Moholkar VS, Goyal A (2014) Screening and optimization of pretreatments for Parthenium hysterophorus as feedstock for alcoholic biofuels. Appl Energy 129:195–206
Harun R, Danquah MK (2011) Enzymatic hydrolysis of microalgal biomass for bioethanol production. Chem Eng J 168:1079–1084
Jodayree S, Smith JC, Tsopmo A (2012) Use of carbohydrase to enhance protein extraction efficiency and antioxidative properties of oat bran protein hydrolysates. Food Res Int 46:69–75
Lan EI, Liao JC (2013) Microbial synthesis of n-butanol, isobutanol, and other higher alcohols from diverse resources. Bioresour Technol 135:339–349
Abou-Shanab RAI, Hwang JH, Cho Y, Min B, Jeon BH (2011) Characterization of microalgal species isolated from fresh water bodies as a potential source for biodiesel production. Appl Energy 88(10):3300–3306
Kanz T, Bold HC (1969) Physiological studies, morphological and taxonomical investigation of Nostoc and Anabaena in culture. Publication No. 6924. University of Texas, Austin
Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356
Ajisaka K, Fujimoto H, Miyasato M (1998) An alpha-L-fucosidase from Penicillium multicolor as a candidate enzyme for the synthesis of a (1->3)-linked fucosyl oligosaccharides by transglycosylation. Carbohyd Res 309:125–129
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275
Barbarino E, Lourenco SO (2005) An evaluation of methods for extraction and quantification of protein from marine macro- and microalgae. J Appl Phycol 17:447–460
Chow PS, Landhausser SM (2004) A method for routine measurements of total sugar and starch content in woody plant tissues. Tree Physiol 24:1129–1136
Huo YX, Cho KM, Rivera JGL, Monte E, Shen CR, Yan YJ, Liao JC (2011) Conversion of proteins into biofuels by engineering nitrogen flux. Nat Biotechnol 29:346–U160
Shen CR, Liao JC (2008) Metabolic engineering of Escherichia coli for 1-butanol and 1-propanol production via the keto-acid pathways. Metab Eng 10:312–320
El-Mashad HM (2015) Biomethane and ethanol production potential of Spirulina platensis algae and enzymatically saccharified switchgrass. Biochem Eng J 93:119–127
Ask M, Olofsson K, Di Felice T, Ruohonen L, Penttila M, Liden G, Olsson L (2012) Challenges in enzymatic hydrolysis and fermentation of pretreated Arundo donax revealed by a comparison between SHF and SSF. Process Biochem 47:1452–1459
Chen R, Yue ZB, Deitz L, Liu Y, Mulbry W, Liao W (2012) Use of an algal hydrolysate to improve enzymatic hydrolysis of lignocellulose. Bioresour Technol 108:149–154
Kumar S, Gupta R, Kumar G, Sahoo D, Kuhad RC (2013) Bioethanol production from Gracilaria verrucosa, a red alga, in a biorefinery approach. Bioresour Technol 135:150–156
Rosgaard L, Andric P, Dam-Johansen K, Pedersen S, Meyer AS (2007) Effects of substrate loading on enzymatic hydrolysis and viscosity of pretreated barley straw. Appl Biochem Biotech 143:27–40
Ingesson H, Zacchi G, Yang B, Esteghlalian AR, Saddler JN (2001) The effect of shaking regime on the rate and extent of enzymatic hydrolysis of cellulose. J Biotechnol 88:177–182
Borines MG, de Leon RL, Cuello JL (2013) Bioethanol production from the macroalgae Sargassum spp. Bioresour Technol 138:22–29
Shuler MLKF (2002) Bioprocess engineering, 2nd edn. Prentice Hall, NJ
Kim KH, Choi IS, Kim HM, Wi SG, Bae HJ (2014) Bioethanol production from the nutrient stress-induced microalga Chlorella vulgaris by enzymatic hydrolysis and immobilized yeast fermentation. Bioresour Technol 153:47–54
Dumay J, Clement N, Morancais M, Fleurence J (2013) Optimization of hydrolysis conditions of Palmaria palmata to enhance R-phycoerythrin extraction. Bioresour Technol 13:21–27
Kermanshahi-Pour A, Sommer TJ, Anastas PT, Zimmerman JB (2014) Enzymatic and acid hydrolysis of Tetraselmis suecica for polysaccharide characterization. Bioresour Technol 173:415–421
Trivedi N, Gupta V, Reddy CRK, Jha B (2013) Enzymatic hydrolysis and production of bioethanol from common macrophytic green alga Ulva fasciata Delile. Bioresour Technol 150:106–112
Lee OK, Kim AL, Seong DH, Lee CG, Jung YT, Lee JW, Lee EY (2013) Chemo-enzymatic saccharification and bioethanol fermentation of lipid-extracted residual biomass of the microalga, Dunaliella tertiolecta. Bioresour Technol 132:197–201
Woods L, Riccobono M, Mehan N, Hestekin J, Beitle R (2011) Synergistic effect of abrasive and sonication for release of carbohydrate and protein from algae. Sep Sci Technol 46:601–604
Tang SH, Hettiarachchy NS, Shellhammer TH (2002) Protein extraction from heat-stabilized defatted rice bran. 1. Physical processing and enzyme treatments. J Agric Food Chem 50:7444–7448
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This work was supported by the Mid-career Researchers Program (the National Research Foundation of Korea, 2013069183).
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Eldalatony, M.M., Kabra, A.N., Hwang, JH. et al. Pretreatment of microalgal biomass for enhanced recovery/extraction of reducing sugars and proteins. Bioprocess Biosyst Eng 39, 95–103 (2016). https://doi.org/10.1007/s00449-015-1493-5
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DOI: https://doi.org/10.1007/s00449-015-1493-5