Electronic Supplementary Materials (ESM) for:
Lipase-Catalyzed Synthesis of Saccharide-Fatty Acid Esters Utilizing
Solvent-Free Suspensions: Effect of Acyl Donors and Acceptors, and
Enzyme Retention Activity
(JAOCS)
Ran Ye, Douglas G. Hayes*
Department of Biosystems Engineering and Soil Science, University of
Tennessee, Knoxville, TN 37996-4531
*To
whom correspondence should be addressed: 2506 E.J. Chapman Dr.,
Knoxville, TN 37996-4531.
Tel: (865) 974-7991, FAX: (865) 974-4514, E-mail address: dhayes1@utk.edu
Figure S1. Effect of acyl acceptor type (given in legend) on the time course of
reaction for immobilized Rhizomucor miehei lipase- (RML-) catalyzed synthesis
of saccharide-oleic acid esters in a packed-bed bioreactor (PBBR)-based
bioreactor system at 53oC that utilizes solvent-free suspensions of saccharide
crystals. Plotted are the a. ester concentration, b. saccharide concentration, and
(inset) liquid-phase water concentration. Bioreactor system and conditions
described in Table 1 of the main paper.
Figure S2. Effect of acyl acceptor type on the time course of reaction for RMLcatalyzed synthesis of saccharide–caprylic acid esters in a packed-bed
bioreactor (PBBR)-based bioreactor system at 53oC that utilizes solvent-free
suspensions of saccharide crystals.
saccharide
concentration,
and
Plotted are a. ester concentration, b.
(inset)
liquid-phase
water
concentration.
Bioreactor system and conditions described in Table 1 of the main paper.
Figure S3.
Effect of acyl acceptor (given in legend) on the time course of
reaction for RML-catalyzed solvent-free synthesis of saccharide–lauric or myristic acid esters in a stirred bioreactor (STBR) at 65oC that utilizes solventfree suspensions of saccharide crystals. Plotted are the a. ester concentration, b.
saccharide
concentration,
and
(inset)
liquid-phase
water
Operating conditions described in Table 1 of the main paper.
concentration.
Optical light microscopy of suspensions before and after centrifugation
Experimental
Suspension droplets (after and/or before centrifugation) were
carefully removed to a slice and covered by a cover-slip gently avoiding the
existence of air bubbles. Afterwards the prepared samples (magnification, X 40)
were probed using a light microscope with integrated 3 mega pixel digital camera
(Micromaster* Inverted digital microscopes with infinity optics, from Fisher
Scientific, Pittsburgh, PA USA).
Figure S4. Images from light microscopy, 40X magnification, of suspensions for
4 different acyl acceptors ( a xylose, b glucose, c sucrose, and d fructose) in
solvent-free media (Fructose-oleic acid esters/ oleic acid, 25/75, w/w) at 25oC
after forming suspensions as mentioned in Table 1 of the main paper, but before
the centrifugation step is invoked.
Figure S5. Images from light microscopy, 40X magnification, of suspensions for
4 different acyl acceptors (a xylose, b glucose, c sucrose, and d fructose) in
solvent-free media (fructose-oleic acid esters/ oleic acid, 25/75, w/w) at 25oC
after completion of centrifugation. Further conditions are given Table 1 of the
main paper.
Figure S6. Images from light microscopy, 40X magnification, of suspensions for
two different acyl acceptors before centrifugation (80oC):
a fructose and b
glucose; and after centrifugation 25oC: c fructose and d glucose) in solvent-free
media (fructose-oleic acid esters / caprylic acid / oleic acid 25 / 72.1 / 2.9 w/w/w).
Operating conditions for the preparation of suspensions in fatty acid equal those
of Fig 4 of the main paper.
Figure S7. Time course of reaction for the solvent-free synthesis of fructose
oleate at 65oC in PBBR-based bioreactor system with in-situ water removal
method (Vacuum+N2 gas bubbling) for determining the operational stability of
immobilized R. miehie lipase (RML)
Plotted are the average and standard
deviation (error bars) for the four successive runs. Further information is given in
Table 3.