Valorization of agro-industrial wastes to produce hydrolytic enzymes by fungal solid-state fermentation

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1 Valrizatin f agr-industrial wastes t prduce hydrlytic enzymes by fungal slid-state fermentatin Cristina Marz, A.B. Díaz, I. Car and A. Blandin University f Cádiz

2 Intrductin DISCARDED Orange peels (ORA) Grape Pmace Exhausted sugar beet csettes (ESBC) Rice Husk 1

3 Intrductin Orange peels (ORA) Exhausted sugar beet csettes (ESBC) Grape Pmace Rice Husk Enzymes Biethanl Lactic acid Etc. 2

4 Intrductin. Slid-State Fermentatin Slid-state fermentatin (SSF) is a prcess where Fungus grws n the slid surface. Requirements: Temperature Misture Nutrients 3

5 Intrductin. Aim f the prject Orange peels (ORA) Enzymatic Hydrlysis Slid-State Fermentatin (SSF) Hydrlytic Enzymes Exhausted sugar beet csettes (ESBC) 4

6 Materials and Methds. Raw material ORA ESBC ORASSF Prcess SSF Hydrlysis SSF Hydrlysis Hydrlysis Slid cnditining Sized Washed Milled Dried Hydrated (3%) Dried Sized Washed Milled Dried SSF Sterilized (120ºC 20 min) ORA: Orange peels ESBC: Exhausted sugar beet cssettes ORASSF: Orange peels after SSF prcess 5

7 Materials and Methds. Slid-state fermentatin SSF Cnditins Grams f Dry Slid 5g r 100g (ORA / ESBC) Temperature 30 ºC Misture 70 % Fungus Aspergillus awamri Inculum spres/g cncentratin Nutrient slutin (g/l) urea (NH 4 ) 2 SO 4 KH 2 PO 4 FeSO 4 7H 2 O ZnSO 4 7H 2 O MgSO 4 7H 2 O CuSO 4 5H 2 O Enzymatic extract cnditins Grams f Dry Slid 5g r 100g (ORA / ESBC) Extractin slvent 0.1 % (v/v) Tween 80 Liquid slid rati 10:1 Temperature 4 ºC Agitatin 150 r.p.m. Time 30 min Centrifugatin 10,000 r.p.m 10 min 6

8 Materials and Methds. Enzymatic hydrlysis Hydrlysis cnditins Grams f Dry Slid Liquid slid rati 6 g (ORA / ESBC / ORASSF) 10:1 / 14:1 Temperature 50 ºC Agitatin 150 r.p.m ph ph 5 (Citrate-phsphate buffer, 0.1M) Exp. Enzymatic ccktails Enzymes Enzymatic activity Exp. 1 Cellulase 1110 FPU/g slid Exp. 2 Ex. 3 Cellulase Xylanase Pectinase β-glucsidase Cellulase Xylanase (SSF) Pectinase (SSF) β-glucsidase (SSF) 1110 FPU/g slid 23.4 U/g slid 31 U/g slid 0.7 U/g slid 1110 FPU/g slid 23.4 U/g slid 31 U/g slid 3.25 U/g slid 7

9 Results and Discussin Slid-State Fermentatin f ORA and ESBC FPase and β-glucsidase activities were negligible Enzyme activities are higher n ESBC Slid Enzyme Maximum activity Day ORA Xylanase 29,600 4 th ORA Pectinase 19,300 2 nd ESBC Xylanase 35,700 8 th ESBC Pectinase 29,700 8 th 8

10 Results and Discussin Slid-State Fermentatin f ORA and ESBC Enzyme Xylanase ex PG Waste ORA ESBC ORA ESBC μ k D k E R Díaz, A., Blandin, A., Webb, C., Car, I.: Mdelling f different enzyme prductins by slid-state fermentatin n several agr-industrial residues. Appl. Micrbil. Bitechnl. 100, , (2016) 9

11 Results and Discussin Slid-State Fermentatin f ORA Extracts btained frm SSF f ORA in rux bttles (100 g f slid) were freeze-dried and their enzymatic activities measured Enzyme activity (U/Kg) Enzyme Xylanase Pectinase Cellulase B Glucsidase 10

12 Results and Discussin. Hydrlysis The highest yield is btained when cellulase is supplemented with enzymatic extract prduced by SSF RS (g/l) Hydrlysis f Exhausted Sugar Beet Cssettes 10 Cmmercial Cellulase (CC) 0 CC sup. with enzymatic extract (SSF) CC sup. with enzymatic cmmercial extract Time (h) 11

13 Results and Discussin. Hydrlysis Hydrlysis f Orange peels after SSF Hydrlysis f Orange peels RS (g/l) RS (g/l) Time (h) Cmmercial Cellulase (CC) CC sup. with enzymatic extract (SSF) CC sup. with enzymatic cmmercial extract Time (h) 12

14 Cnclusins Orange peels and exhausted sugar beet cssettes are gd natural media fr the grwth f Aspergillus awamri by SSF and fr the prductin f hydrlytic enzymes The enzymatic extract btained by SSF n bth wastes is cmpsed f a brad range f hydrlytic enzymes, mainly xylanases and pectinases The enzymatic extract btained by SSF can be used as a supplement ccktail f cellulase fr saccharificatin prcesses f ligncellulsic material Enzymatic extract btained frm SSF n range peels prvides higher hydrlysis yield fr all residues tested than a cmmercial enzymatic ccktail with the same enzymatic activities 13

15 Valrizatin f agr-industrial wastes t prduce hydrlytic enzymes by fungal slid-state fermentatin Cristina Marz, A.B. Díaz, I. Car and A. Blandin University f Cádiz cristina.marz@uca.es

PRODUCTION OF VALUE ADDED FOOD DERIVED SUB PRODUCTS VIA SSF USING Rhizopus sp.

PRODUCTION OF VALUE ADDED FOOD DERIVED SUB PRODUCTS VIA SSF USING Rhizpus sp. Jne Ibarruri, AZTI (jibarruri@azti.es) Fd Research Unit. Efficient and Sustainable Prcesses Intrductin Brewer s spent grain