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

Transcription

1 PRODUCTION OF VALUE ADDED FOOD DERIVED SUB PRODUCTS VIA SSF USING Rhizpus sp. Jne Ibarruri, AZTI Fd Research Unit. Efficient and Sustainable Prcesses

2 Intrductin Brewer s spent grain (BSG) 85 % f ttal by prducts generated during beer prductin > 7 millin tnes f residues in Eurpe breweries annually Rich in fiber (70 %) and prtein (20 %) enzymes Antixidant peptides Antihypertensive peptides Phenlic cmpunds High ecnmical cst Value Value Micrbial fermentatin 1

3 Intrductin SSF as valrizatin strategy Fungi main micrrganisms used in SSF Knwn t prduce enzymes which degrade plant cell walls Imprve bichemical cmpsitin and biactivity f the emplyed substrates Micrrganism fr SCP prductin shuld have High affinity fr the substrate Lw nutritinal requirements Ability t use cmplex substrates Ability t develp high cell density Gd tlerance t temperature and ph Balanced prtein and lipid cmpsitin Lw nucleic acid cntent Gd digestibility and nn txic 2

4 Intrductin SSF as valrizatin strategy Rhizpus sp. ne f the mst prmising fungal genus Cnsume a great range f carbn surces Enzyme and rganic acid prductin By prduct valrizatin Advantages fr feed applicatin Generally recgnized as safe (GRAS) by the FDA Inculum fr human fd prductin (tempeh, ragi ) Significantly reduced the amunt f testing required High cntent f prtein (35 60 %) Lw levels f nucleic acids Pleasant taste and smell f the fungal bimass 3

5 Materials and Methds Analytical methds Fd derived Rhizpus strain islated and characterized in ur labratry (ROR004) [1] and a strain derived frm ROR004 after mutagenesis experiments (ROR00G), UV, Ethidium brmide and MNNG. Dry weight: drying at 60 ºC, until cnstant weight Prtein and ttal nitrgen cntent: Kjeldahl methd Ttal Amin acids:hydrlysiswith6nhclandhplc Essential Amin Acid Index: (2) 1 1 (1) Ibarruri, J., Hernández, I.: Rhizpus ryzae as fermentatin agent in fd derived subprducts. Waste and Bimass Valrizatin (2017). di: /s (2) Cncentratin and characterizatin f micralgae prteins frm Chlrella pyrenidsa Ashish G. Waghmare, Manj K. Salve, Jean Guy LeBlanc and Shalini S. Arya. Biresurces and biprcessing (2016) 3:16 (2) Antixidant activity: DPPH radical scavenging activity (TEAC, Trlx equivalent capacity) Ttal phenlic cntent (TPC): Flin Cicalteu methd (GAE, Gallic acid equivalent) Reducing Sugar: Dinitrsalicylic acid (DNS) methd Sluble prtein : BSA methd Degree f hydrlysis (DH): Phthaldialdehyde (OPA) methd Antibacterial activity: agar diffusin methd Salmnella enterica (CECT 4156) and Escherichia cli (CECT 516) 4

6 Materials and Methds BSG SSF Prvided by Bga Cperative (an artisanal brewery situated in Mungia, Spain) Sterilized at 110 ºC, 15 min Inculatin (10 6 cfu/g) Density was maintained at 0.09 g/cm 2 Sterile Petri plate (surface area cm 2 ), 30 ºC, frm T0 t T192 h t determine the variables that change alng the SSF Factrial design (2 3 ) (8 runs) and central pints (4 runs) was used fr ptimal cnditins determinatin Factr Lw( ) 0 High (+) A: Time (h) B: Temperature (ºC) C: Strain 1 (Mutant) 2 (ROR004) 5

7 Results Crude prtein (%DM) Fermentatin time (h) Sluble prtein (mg/g DM) Antibacterial activity was nt detected in the extracts btained frm fermented BSG TPC (µg GAE/g DM) TEAC (µg Trlx/g DM) Fermentatin time (h) DH (%) DPPH reductin (%) Nt differences in reducing sugars Fermentatin time (h) 6

8 Results SSF EVOLUTION Time 0 h Time 72 h Time 120 h Ideal substrate fr SSF cmpsitin particle size Time 144 h Time 168 h Time 192 h Small particles with rigid internal structureallwacmpletefungal grwth and a cmplete use f the inter particle space Hmgenus fermented material andavidthepresencefunderfermented areas 7

9 Results SSF ptimizatin Mdel P value R 2 Adjusted Observed Prtein * 65.4 Sluble prtein * 96.8 DH * 77.7 TEAC * 78.4 DPPH * 82.9 TPC * 92.6 Prtein (% DM) ±7.6 Sluble prtein (mg/g DM) ± 3.8 DH (%) ± 7.0 TEAC (µg TE/g DM) ± 102 DPPH reductin (%) ± 1.9 TPC (µg GAE/g DM) ± 112 Overall strategy fr BSG valrizatin is t ptimize all respnses at a time 30 ºC, 216 h f fermentatin time 8

10 Results Amin Acid Prfile (3) FAO/WHO: Prtein and amin acid requirements in human nutritin. Expert Cnsultatin, vl WHO Technical Reprt Series, pp Wrld Health Organizatin, Geneva, (2007). (4) FAO (WHO) (1974) Prtein advisry grup guidelines n. 15 n the nutritinal and safety aspects f nvel surces f prtein fr animal feeding. United Natins, Rme EAA prfile (%) EAA cntent (% DM BSG) Unfermented BSG Fermented BSG Unfermented BSG Fermented BSG His 2.9 ±0.1 a 2.7 ±0.5 a 0.6 ±0.0 a 0.8 ±0.1 b Ile 3.7 ±0.2 a 3.7 ±0.2 a 0.7 ±0.0 a 1.2 ±0.1 b Leu 8.2 ±0.1 a 7.6 ±0.3 a 1.7 ±0.0 a 2.4 ±0.1 b Lys 6.6 ±0.2 a 6.8 ±0.4 a 1.3 ±0.0 a 2.1 ±0.1 b Met 1.1 ±0.0 a 1.6 ±0.2 b 0.2 ±0.0 a 0.5 ±0.1 b Cys 0.9 ±0.2 a 1.3 ±0.3 a 0.2 ±0.1 a 0.4 ±0.1 b Phe 5.5 ±0.0 a 4.7 ±0.4 a 1.1 ±0.0 a 1.5 ±0.1 b Tyr 3.7 ±0.2 a 4.4 ±1.1 a 0.7 ±0.0 a 1.4 ±0.3 b Thr 4.0 ±0.3 a 4.6 ±0.3 a 0.8 ±0.1 a 1.4 ±0.1 b Val 5.8 ±0.2 a 5.9 ±0.5 a 1.2 ±0.0 a 1.8 ±0.1 b Ttal essential 42.4 ±0.2 a 43.1 ±2.3 a 8.6 ±0.0 a 13.6 ±0.7 b 9

11 Results Amin Acid Prfile (3) FAO/WHO: Prtein and amin acid requirements in human nutritin. Expert Cnsultatin, vl WHO Technical Reprt Series, pp Wrld Health Organizatin, Geneva, (2007). (4) FAO (WHO) (1974) Prtein advisry grup guidelines n. 15 n the nutritinal and safety aspects f nvel surces f prtein fr animal feeding. United Natins, Rme Fermented BSG Sya prtein Casein FAO Reference FAO Reference His 2.7 ± Thr 4.6 ± Cys+Met 2.9 ± Val 5.9 ± Lys 6.8 ± Ile 3.7 ± Leu 7.6 ± Phe+tyrsine 9.1 ± EAAI FAO (3) 1.58 ± EAAI FAO (4) 1.28 ±

12 Cnclusins Using filamentus fungi as bi fermentatin f BSG leads t a prtein rich substrate with higher essential amin acid cntent and hydrlysis degree which culd imprve prtein digestibility SSF als increases the antixidant activity f the fermented BSG related t the release f phenlic cmpunds SSF is a prmising alternative t revalrize this agr industrial by prduct as ingredient fr feed and fd applicatins BSG is als knwn fr cntaining peptides related t antihypertensive activity and fr specific phenlic cmpunds (ferulic, caffeic and p cumaric acid) related t anti cancer, anti athergenic and anti inflammatry effects Further research is needed t evaluate the effect f SSF by Rhizpus sp. in the liberatin f thse specific peptides and phenlic cmpunds 10

13 Cnclusins Onging research Scaling up in drum bireactr SSF effect f antihypertensive peptide liberatin (psitive preliminary results) Obtained peptides prfile Anti inflammatry and immunstimulatry effects f the fermented prduct Enzyme prductin Ecnmical, nutritinal, safety and envirnmental viability f the prcess Unfermented BSG Fermenetd BSG (30 ºC, 9 days) 11

14 Índice Thank yu fr yur attentin Txatxarramendi ugarte z/g Sukarrieta, Bizkaia Herrera Kaia. Prtualdea z/g Pasaia, Gipuzka Astnd Bidea, Edifici 609 Parque Tecnlógic de Bizkaia Deri, Bizkaia