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Interactions between fungal growth, substrate utilization, and enzyme production during solid substrate cultivation of Phanerochaete chrysosporium on cotton stalks

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Abstract

Fungal pretreatment, using lignin-degrading microorganisms to improve lignocellulosic feedstocks with minimal energy input, is a potential alternative to physiochemical pretreatment methods. Identifying the kinetics for fungal pretreatment during solid substrate cultivation is needed to help establish the processing conditions for effective scale up of this technology. In this study, a set of mathematical models were proposed for describing the interactions between holocellulose consumption, lignin degradation, cellulase, ligninolytic enzyme, and the growth of Phanerochaete chrysosporium during a 14 day fungal pretreatment process. Model parameters were estimated and validated by the System Biology Toolbox in MatLab. Developed models provided sufficiently accurate predictions for fungal growth (R 2 = 0.97), holocellulose consumption (R 2 = 0.97), lignin degradation (R 2 = 0.93) and ligninolytic enzyme production (R 2 = 0.92), and fair prediction for cellulase production (R 2 = 0.61). The models provide valuable information for understanding the interactive mechanisms in biological systems as well as for fungal pretreatment process scale up and improvement.

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Abbreviations

α C :

Growth-associated constant for cellulase formation, (g enzyme protein) (g DM)−1

α L :

Growth-associated constant for ligninolytic enzyme formation, (g enzyme protein) (g DM)−1

β C :

Non-growth-associated constant for cellulase formation, (g enzyme protein) (g DM)−1

β L :

Non-growth-associated constant for ligninolytic enzyme formation, (g enzyme protein) (g DM)−1

μ max :

The maximum specific growth rate, day−1

C FPU :

Cellulase activity per ml of enzyme extract, IU ml−1

C TKN,E :

Amount of TKN per ml of enzyme extract, (g TKN) ml−1

C TKN,base :

Base TKN content in autoclaved cotton stalk extract, (g TKN) ml−1

C TKN,S :

Amount of TKN in pretreated solids, (g TKN) (g DM)−1

CCP:

Cumulative CO2 production (mmol)

COU:

Cumulative oxygen uptake (mmol)

CPR:

CO2 production rate (mmol day−1)

DM:

Dry matter

DML:

Dry matter loss

k E = 1/1070:

Conversion coefficient from cellulase activity to cellulase proteins, (g cellulase proteins) IU−1

k LD :

Coefficient of lignin degradation

k LL :

Coefficient of ligninolytic enzyme production, (g enzyme protein) (g fungal biomass)−1 day−1

k T = 10.3:

Conversion coefficient from TKN to fungal biomass, based on the amount of TKN in pure P. chrysosporium fungal biomass, (g fungal biomass) (g TKN)−1

m :

Maintenance coefficient, (g cellulose and hemicellulose) (g fungal biomass)−1 day−1

m O :

Maintenance coefficient, (mmol O2) (g fungal biomass)−1 day−1

OU:

Oxygen uptake, mmol

OU0 :

Initial oxygen uptake, mmol

OUR:

Oxygen uptake rate, (mmol O2) day−1

P C :

Cellulase concentration, (g enzyme protein) (g DM)−1

P C,0 :

Initial cellulase concentration at time 0, (g enzyme protein) (g DM)−1

P L :

Ligninolytic enzymes concentration, (g enzyme protein) (g DM)−1

P L,0 :

Initial ligninolytic enzymes concentration, (g enzyme protein) (g DM)−1

S C :

Cellulose and hemicellulose content, g (g DM)−1

S C,0 :

Initial cellulose and hemicellulose content, g (g DM)−1

S L :

Lignin content, g (g DM)−1

S L,0 :

Initial lignin content, g (g DM)−1

t :

Pretreatment time, day

TKN:

Total Kjeldahl Nitrogen, %

V = 20:

Volume of enzyme extracts, ml

w :

Dry weight of pretreated sample, g DM

w 0 :

The initial sample dry weight before pretreatment, g DM

X :

The fungal biomass content, g (g DM)−1

X 0 :

Initial fungal biomass content, g (g DM)−1

X max :

Maximum biomass content, g (g DM)−1

Y X/O :

The oxygen to fungal biomass yield coefficient (g−X mmol−1 O2)

Y X/S :

The substrate to fungal biomass yield coefficient, (g fungal biomass) (g cellulose and hemicellulose DM)−1

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Acknowledgments

The authors would like to thank Dr. Van Den Truong (Dept of Food Science, NCSU), Roger Thompson (USDA-ARS, NCSU) and Rachel S. Huie (Dept of Biological and Agricultural Engineering, NCSU) for generously helping with TKN and O2/CO2 analysis.

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Authors and Affiliations

  1. Department of Biological and Agricultural Engineering, North Carolina State University, 277 Weaver Labs, Campus Box 7625, Raleigh, NC, 27695-7625, USA

    Jian Shi, Mari S. Chinn & Ratna R. Sharma-Shivappa

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  1. Jian Shi
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  2. Mari S. Chinn
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Correspondence to Mari S. Chinn.

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Shi, J., Chinn, M.S. & Sharma-Shivappa, R.R. Interactions between fungal growth, substrate utilization, and enzyme production during solid substrate cultivation of Phanerochaete chrysosporium on cotton stalks. Bioprocess Biosyst Eng 37, 2463–2473 (2014). https://doi.org/10.1007/s00449-014-1224-3

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