Måling og styring av hydrolyseprosesser skreddersydd proteinproduksjon

Transcription

1 Måling og styring av hydrolyseprosesser skreddersydd proteinproduksjon Nils Kristian Afseth, Sileshi Wubshet, Ingrid Måge, Ulrike Böcker, Jens Petter Wold, Kenneth Kristoffersen and Diana Lindberg

2 Enzymatic protein hydrolysis in agri-food valorization Enzymatic hydrolysis Lipids Protein hydrolysate Unsoluble proteins / sediments

3 Enzymatic protein hydrolysis in agri-food valorization Bioactive peptides Cosmetics Functional food Pet food Enzymatic hydrolysis Food Feed Oil Sediments for feed

4 Enzymatic hydrolysis (the simple case) Single protein digestion: -Myoglobin (trypsin) LCMS Chromatogram

5 Enzymatic hydrolysis (the real case) LCMS chromatogram of Hydrolysate from MDC Wubshet, S. G. et al. Unpublished data, 2016

6 Bioanalytics and protein characterisation Degree of hydrolysis OPA / TNBS etc. Size distributions size exclusion chromatography Detailed characterization LCMS / NMR

7 Rapid and non-destructive spectroscopic measurements NIR spectroscopy FTIR spectroscopy Raman spectroscopy

8 FTIR provides a biochemical fingerprint Qualitative and quantitative information on protein structure

9 Monitoring enzymatic protein FTIR is sensitive to the amide backbone and the terminal carboxylates hydrolysis - FTIR

10 Monitoring enzymatic protein hydrolysis-ftir (2)

11 Apparent degree of hydrolysis (DH%) FTIR for monitoring enzymatic protein hydrolysis Time FTIR spectra

12 FTIR for monitoring enzymatic protein hydrolysis Chicken muscle Salmon muscle Salmon heads Chicken by-products Salmon backs FTIR spectra - Chicken by-products Ref: Böcker, U., Wubshet, I., Lindberg, D., and Afseth, N.K. Fourier-transform Infrared spectroscopy for qualitative characterization of protein chain reductions in enzymatic hydrolysis, submitted manuscript.

13 Weight Average Molecular Weight (g/mol) Quantitative monitoring of enzymatic protein hydrolysis Average MW vs Hydrolysis time Size exclusion chromatography Hydrolysis time (min.) KF_04_01 KF_04_02

14 Quantitative monitoring of enzymatic protein hydrolysis Size exclusion chromatography FTIR spectroscopy Refs: Wubshet, S., Måge, I., Bocker, U., Lindberg, D., Knutsen, S.H., Rieder, A., Airado-Rodriguez, D. & Afseth, N.K.. FTIR as a rapid tool for monitoring enzymatic protein hydrolysis of food processing by-products, submitted manuscript.

15 A spectroscopic platform for control and optimisation of biotech processes Projects: LiqIR Project leader: Prediktor ( ) - Funded by the Regional research fund Oslofjordfondet, with Biomega, Nutrimar, VEAS, and SINTEF ICT

16 Prediction of hydrolysis outcome based on raw material analysis

17 Prediction of hydrolysis outcome the chicken and turkey case 32 samples of chicken and turkey rest raw materials were collected at Nortura Hærland, spanning variation in chemical composition

18 Raw material variation and quality Process Analytical Technology (PAT): A strategy towards a stable product quality Protein 0.7 Fat Ash 1.2 Moisture 2.1

19 Hydrolysis time AMW Can we predict product quality from raw material and processing settings? Raw material property Process setting Product quality Protein, Ash, Moisture and Fat +

20 Can we predict product quality from raw material and processing settings? Multiblock regression (SO-PLS) Hydrolysis time AMW Spectra -measurement on rawmaterial + Raman Time + 3 comp.

21 Future protein hydrolysis raw material variation can be handled using spectroscopy Raw material properties measured by spectroscopy Raman Process settings Hydrolysis time NIR / FTIR End-product quality Protein yield Average Molecular Weight Refs: Wubshet, S. et al., Spectroscopic process monitoring in enzymatic protein hydrolysis of byproducts: A strategy for stable product quality, manuscript.

22 Concluding remarks Controlling and handling rest raw material complexity and variation is essential for future valorisation Industrial analytical tools are key factors in producing protein products of defiend qualities (e.g. sensorial, functional or bioactive properties) for higher paying markets: Flexible processing for dealing with rest raw material heterogeneity and variations (e.g. depending on season or pre-production steps) Possibilities for real-time process adjustments

23 PepFishing (July 2017-June 2020) A novel combination of FTIR-based screening and ligand fishing technologies for facilitated discovery of antidiabetic peptides from protein hydrolysates.

24 Acknowledgements Industry collaboration: Norilia, Nortura, Biomega and Nutrimar Research collaboration: SINTEF FA and SINTEF ICT Funding from The Norwegian Research Council, Oslofjordfondet, and the foundation for Research levy on Agricultural Products: «HydrolyseMonitor», led by Nofima «Chickenlysis», led by Nortura «LiqIR», led by Prediktor «Cycle», led by Sintef FA «Food quality imaging», led by Nofima