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Application of whey retentate as complex nitrogen source for growth of the polyhydroxyalkanoate producer Hydrogenophaga pseudoflava strain DSM1023

Martin Koller
Martin Koller
, 
Paula Hesse
Paula Hesse
 and 
Gerhart Braunegg
Gerhart Braunegg
   | Apr 24, 2019
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Article Category: Research Article
Published Online: Apr 24, 2019
Page range: 78 - 89
DOI: https://doi.org/10.2478/ebtj-2019-0009
Keywords
© 2019 Martin Koller, Paula Hesse, Gerhart Braunegg, published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Figure 1

Processing of whey towards a feedstock for PHA production (based on (34)).
Processing of whey towards a feedstock for PHA production (based on (34)).

Figure 2

Time patterns of OD (a), CDM (b), and inorganic nitrogen source ammonium sulfate (c). H. pseudoflava cultivated on shaking flask scale for 24 h on hydrolyzed whey permeate as carbon source and ammonium sulfate as inorganic nitrogen sources; comparison of growth patterns with addition of different complex nitrogen sources (yeast extract, CSL, casein hydrolysate, malt extract, pressed grass silage juice, meat extract, and not hydrolyzed whey retentate).
Time patterns of OD (a), CDM (b), and inorganic nitrogen source ammonium sulfate (c). H. pseudoflava cultivated on shaking flask scale for 24 h on hydrolyzed whey permeate as carbon source and ammonium sulfate as inorganic nitrogen sources; comparison of growth patterns with addition of different complex nitrogen sources (yeast extract, CSL, casein hydrolysate, malt extract, pressed grass silage juice, meat extract, and not hydrolyzed whey retentate).

Figure 3

(a) times curves of OD (420 nm) during the phase of microbial growth (t = 0-15 h); (b): CDM at time 0 h (black bars), 11.75 h (grey bars), and 26.25 h (white bars); A: only inorganic N-source, B: 0.5 g/L HWR, C: 2 g/L HWR, D: 5 g/L HWR, E: 0.5 g/L NHWR, F: 2 g/L NHWR, G: 5 g/L NHWR. H. pseudoflava on hydrolyzed whey permeate as carbon source and different additions of HWR or NHWR. Error bars indicate the deviations between two parallel cultivations.
(a) times curves of OD (420 nm) during the phase of microbial growth (t = 0-15 h); (b): CDM at time 0 h (black bars), 11.75 h (grey bars), and 26.25 h (white bars); A: only inorganic N-source, B: 0.5 g/L HWR, C: 2 g/L HWR, D: 5 g/L HWR, E: 0.5 g/L NHWR, F: 2 g/L NHWR, G: 5 g/L NHWR. H. pseudoflava on hydrolyzed whey permeate as carbon source and different additions of HWR or NHWR. Error bars indicate the deviations between two parallel cultivations.

Figure 4

Remaining concentrations of ammonium sulfate (a), total sugars (sum glucose + galactose, b), glucose (c), and galactose at time 0 h (black bars), 11.75 h (grey bars), and 26.25 h (white bars). A: only inorganic N-source, B: 0.5 g/L HWR, C: 2 g/L HWR, D: 5 g/L HWR, E: 0.5 g/L NHWR, F: 2 g/L NHWR, G: 5 g/L NHWR. H. pseudoflava on hydrolyzed whey permeate as carbon source and different additions of HWR or NHWR. Error bars indicate the deviations between two parallel cultivations.
Remaining concentrations of ammonium sulfate (a), total sugars (sum glucose + galactose, b), glucose (c), and galactose at time 0 h (black bars), 11.75 h (grey bars), and 26.25 h (white bars). A: only inorganic N-source, B: 0.5 g/L HWR, C: 2 g/L HWR, D: 5 g/L HWR, E: 0.5 g/L NHWR, F: 2 g/L NHWR, G: 5 g/L NHWR. H. pseudoflava on hydrolyzed whey permeate as carbon source and different additions of HWR or NHWR. Error bars indicate the deviations between two parallel cultivations.

Figure 5

Concentrations of PHA (a), residual biomass (CDM minus PHA, b), mass fraction m of PHA in CDM (c), and volumetric PHA productivity at time 0 h (black bars), 11.75 h (grey bars), and 26.25 h (white bars). A: only inorganic N-source, B: 0.5 g/L HWR, C: 2 g/L HWR, D: 5 g/L HWR, E: 0.5 g/L NHWR, F: 2 g/L NHWR, G: 5 g/L NHWR. H. pseudoflava on hydrolyzed whey permeate as carbon source and different additions of HWR or NHWR. Error bars indicate the deviations between two parallel cultivations.
Concentrations of PHA (a), residual biomass (CDM minus PHA, b), mass fraction m of PHA in CDM (c), and volumetric PHA productivity at time 0 h (black bars), 11.75 h (grey bars), and 26.25 h (white bars). A: only inorganic N-source, B: 0.5 g/L HWR, C: 2 g/L HWR, D: 5 g/L HWR, E: 0.5 g/L NHWR, F: 2 g/L NHWR, G: 5 g/L NHWR. H. pseudoflava on hydrolyzed whey permeate as carbon source and different additions of HWR or NHWR. Error bars indicate the deviations between two parallel cultivations.

Figure 6

Growth rates rX and specific growth rates μ for time interval between t = 5 and 8.75 h (a and b, respectively), and for time interval between t = 5 and 11.75 h (c and d, respectively). H. pseudoflava on hydrolyzed whey permeate as carbon source and different additions of HWR or NHWR. Full lines with full symbols: NHWR; dashed lines with open symbols: HWR. rX = dOD/dt; μ = rX/OD
Growth rates rX and specific growth rates μ for time interval between t = 5 and 8.75 h (a and b, respectively), and for time interval between t = 5 and 11.75 h (c and d, respectively). H. pseudoflava on hydrolyzed whey permeate as carbon source and different additions of HWR or NHWR. Full lines with full symbols: NHWR; dashed lines with open symbols: HWR. rX = dOD/dt; μ = rX/OD

Figure 7

a) Dixon plot to determine inhibition constant Ki for HWR on growth of H. pseudoflava; b) Monod kinetics, c) Haldane kinetics for utilization of NHWR. H. pseudoflava on hydrolyzed whey permeate as carbon source and different additions of HWR H. pseudoflava on hydrolyzed whey permeate as carbon source and different additions of HWR or NHWR.
a) Dixon plot to determine inhibition constant Ki for HWR on growth of H. pseudoflava; b) Monod kinetics, c) Haldane kinetics for utilization of NHWR. H. pseudoflava on hydrolyzed whey permeate as carbon source and different additions of HWR H. pseudoflava on hydrolyzed whey permeate as carbon source and different additions of HWR or NHWR.

Chemical composition of milk and different types of whey (based on (33))

Compound (% (w/w))MilkSweet WheyDry Sweet whey (powder)Fermented whey (powder)Dry sweet whey (powder)Whey Permeate (this study)Whey Retentate (this study)
Lactoseca. 4.84.7–4.973.54.5–4.965.62314
Lactic acid-tracestraces0.57n.d.n.d.
Proteins3.4-3.50.75–1.112.90.4512.30.7513
Lipids>4.20.15–0.21.1traces1.0n.d.3-4
Minerals(e.g., calcium, phosphate)~0.7ca. 0.780.6-0.711ca. 2.7ca. 0.7

Kinetic constants for whey retentate for H. pseudoflava in a medium with 2 g/L ammonium sulfate as inorganic nitrogen source established according to Haldane and Monod

EquationKsμmaxKi
Haldane0.180.4614.98
Monod0.110.42-

Growth rates (rX) and specific growth rates (μ) of cultures from the exponential growth phase expressed based on OD values

TimeOnly inorg. N-sourceYeast extractMalt extractMeat extractCSLCasein HydrolysatePressed grass silage juiceWhey retentate
rX (dOD/dt)5-8 h0.521.050.700.940.630.600.310.44
rX (dOD/dt)8-11 h1.111.881.641.841.741.581.531.68
μ (1/h)5-8 h0.220.340.260.310.200.240.090.16
μ (1/h)8-11 h0.280.300.340.310.340.370.350.42
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