Lactobacillus casei as a biocatalyst for biofuel production
Elena Vinay-Lara1, Song Wang2, Lina Bai3, Ekkarat Phrommao1, Jeff R. Broadbent 4, and James L. Steele1*
1Department of Food Science, University of Wisconsin-Madison, WI 53706, USA
2Collegeof Food Science, Northeast Agricultural University, Harbin, Heilongjiang Province, China.
3College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, Hubei Province 430070, China
4 Department of Nutrition, Dietetics, and Food Sciences, Utah State University UT 84322-8700, USA
1 Corresponding Author: Mailing address: 127B Babcock Hall, 1605 Linden Drive, Madison, WI 53706. Phone: (608) 262-5960. Fax: (608) 262-6872. Email:
Online Resources
Online Resource 1. Composition and concentration of modified synthetic corn stover hydrolysate base medium (mSynH)
Materials / Concentration (mM)Glucose / 50
KH2PO4 / 3.4375
K2HPO4 / 6.5625
(NH4)2SO4 / 30
KCl / 45
NaCl / 25
CaCl2·6H2O / 0.09
MgCl2. 6H2O / 1
MnCl2*4H20 / 0.1
ZnCl2 / 0.012
CuCl2 / 0.00001
CoCl2·6H2O / 0.00003
(NH4)6Mo7O24•4H2O / 0.000003
FeCl3·6H2O / 0.017
H3BO3 / 0.01
2,3 -Dihydroxybenzoic acid / 0.01
p-Hydroxybenzoic acid / 0.01
Sodium formate / 10
Sodium nitrate / 0.1
Sodium succinate / 0.5
Glycerol / 3
L-lactic acid / 38
Racemic (L,D-lactic acid) / 2
Betaine.H2O / 0.7
Choline Chloride / 0.7
DL-Carnitine / 0.2
Sodium pyruvate / 30
Sodium citrate / 10
DL-Malic acid / 10
Thiamine HCl / 0.01
Calcium Pantothenate / 0.01
p-aminobenzoic acid / 0.01
*D-Biotin / 0.0008
*Folic Acid / 0.0022
*Niacinamide / 0.0081
*Pyridoxine hydrochloride / 0.0048
*Riboflavin / 0.0005
*Pyridoxal HCl / 0.0598
AA Stock solution#1(50X) / 5
AA Stock solution#2(10X)
Tween 80 / 0.1%
*Compounds added to allow Lactobacillus casei growth.
Online Resource 2. Lignotoxic compounds used in the study.
Compound / Concentration (mM)Furan derivatives
Hydroxymethylfurfural (HMF) / 0.55
Phenolic compounds
Feruloyl amide / 2.75
Coumaroyl amide / 2.75
ρ – Coumaric acid / 1.05
Ferulic acid / 0.36
Benzoic acid / 0.48
Syringic acid / 0.08
Cinnamic acid / 0.09
Vanillic acid / 0.09
Caffeic acid / 0.01
Vanillin / 0.13
Syringaldehyde / 0.16
4- Hydroxybenzeldehyde / 0.2
4- Hydroxyacetophenone / 0.02
Online Resource 3. Ethanol and isobutanol concentrations (v/v %) needed to reduce maximum growth rate (µmax) and optical density (ODmax) by 50%.
Ethanol (v/v %) / Isobutanol (v/v %)umax / ODmax / umax / ODmax
L. casei E1 / 4.5b / ± / 0.08 / 6.8a / ± / 0.07 / 1.3a / ± / 0.02 / 1.3a / ± / 0.02
E. coli GLBRC E1 / 5.1a / ± / 0.46 / 1.9b / ± / 0.01 / 0.8b / ± / 0.06 / 0.4b / ± / 0.01
Z. mobilis 31821 / 3.8c / ± / 0.20 / 1.9b / ± / 0.03 / 0.8b / ± / 0.05 / 0.4b / ± / 0.01
Online Resource 4. Maximum growth rate (µmax) and optical density (ODmax) exhibited by the three ethanologen strains, during growth in modified synthetic corn stover hydrolysate and mSynH supplemented with different stressors. Conditions include osmotic stress (O), Acetate (A) and lignotoxin compounds (L) O, A and L, and combinations of these stressors. Conditions that share at least one superscript are not statistically different. First superscript letters are to be compared within columns, with mSynH condition as control. Second superscript letters are to be compared between ethanologens.
L. casei E1 / E. coli GLBRC E1 / Z. mobilis ATCC 31821Condition / μmax / ODmax / μmax / ODmax / μmax / ODmax
mSynH / 0.18±0.01a / 1.84±0.01a / 0.13±0.01a / 2.11±0.01a / 0.12±0.01a / 1.75±0.01a
O / 0.14±0.01c / 1.64±0.01b / 0.11±0.01b / 1.91±0.01a / 0.10±0.01b / 1.58±0.01ab
A / 0.17±0.01b / 1.74±0.01b / 0.07±0.01d / 1.78±0.01b / 0.07±0.01c / 1.74±0.01a
L / 0.12±0.01d / 1.69±0.01b / 0.08±0.01c / 1.20±0.01d / 0.06±0.01d / 1.03±0.01d
OA / 0.13±0.00c / 1.62±0.01c / 0.11±0.01b / 1.48±0.01b / 0.10±0.00b / 1.50±0.01b
OL / 0.10±0.01e / 1.32±0.01e / 0.07±0.00d / 1.36±0.01c / 0.07±0.01cd / 1.37±0.01c
AL / 0.11±0.01d / 1.51±0.01d / 0.04±0.01e / 0.55±0.01f / 0.04±0.01e / 0.46±0.01f
OAL / 0.10±0.01e;a / 1.23±0.01f / 0.03±0.01f;b / 0.66±0.01e / 0.04±0.01e;b / 0.79±0.01e
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