'Clean' hydrolase reactions using commercial washing powder

Article abstract of DOI:10.1039/c9ra05828a

We report the use of commercial laundry powder as a biocatalyst for a range of lipase-catalysed reactions including (trans)esterification, ester hydrolysis and chemoenzymatic epoxidation reactions. The enzymatic laundry powder exhibited excellent stability and recyclability, making it a readily available and cheap biocatalyst for chemical transformations.

Full text of DOI:10.1039/c9ra05828a

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‘Clean’ hydrolase reactions using commercial
washing powder†
Cite this: RSC Adv., 2019, 9, 24039
b
b
b
Jie Zhang,a Fabio Tonin,
Wuyuan Zhang,
Peter-Leon Hagedoorn,
Lloyd Malleeb and Frank Hollmann
b
*
´
Received 27th July 2019
Accepted 29th July 2019
We report the use of commercial laundry powder as a biocatalyst for a range of lipase-catalysed reactions
including (trans)esterification, ester hydrolysis and chemoenzymatic epoxidation reactions. The enzymatic
laundry powder exhibited excellent stability and recyclability, making it a readily available and cheap
biocatalyst for chemical transformations.
DOI: 10.1039/c9ra05828a
rsc.li/rsc-advances
Modern laundry powders are complex compositions consisting
As a rst model reaction we chose the transesterication
of much more than just surfactants containing more than 25 between ethyl acetate with isopentyl alcohol (Fig. 1). For this,
different ingredients.1 Amongst them, enzymes are frequently the ELP was suspended in a mixture of both reagents in the
included. Enzymes increase the washing performance by absence of any further solvent. It is worth mentioning here that
degrading poorly water soluble polymeric carbohydrates, performing this (and all subsequent reactions) in the absence of
proteins and triglycerides. Particularly hydrolytic enzymes such ELP or using thermally treated ELP (see ESI† for details on the
as cellulases, amylases, proteases and lipases are found in so- thermal enzyme inactivation procedure) gave trace amounts of
called enzymatic laundry powders (ELPs).2 Having this in product only. Therefore, we can assume that all reactions re-
mind, ELP may also be considered as a cheap and readily ported here are indeed enzymatic reactions.
available source of biocatalysts. Hence, they may be considered
A preliminary characterisation of the parameters inuencing
as affordable catalysts e.g. for education purposes or for the ELP-catalysed transesterication reaction revealed that the
research groups with limited nancial possibilities. In fact, system is very well behaved. Increasing the concentration of the
ELPs have been used to isolate DNA from tissue,3 gels,4 blood,5 acyl donor positively inuenced the product formation rate
or hair.6
(Fig. 1a). The same is true for the catalyst loading (Fig. 1b). The
Preparative applications of ELPs, however, have so far not reaction rate steadily increased with increasing ELP concen-
been considered. In this respect, especially the lipases con- trations. Also the inuence of the reaction temperature on the
tained in ELPs represent an interesting starting point. Today, productivity of the transesterication reaction was in line with
lipases are established catalysts for the synthesis of a broad our expectations exhibiting an optimal value at 60 ꢀC, which is
range of compounds ranging from chiral alcohols and amines also the laundry temperature recommended by the producer.
through kinetic resolution of racemic starting materials7 to
Quite interestingly, the method of physical mixing the
cosmetic esters (Scheme 1).8 Also so-called perhydrolysis reac- reaction mixture had a signicant effect on the reaction rate and
tions yielding peracids to mediate chemical oxidation reactions
are gaining increasing attention (Scheme 1).9,10 We therefore set
out to explore the use of ELP to catalyse some typical lipase
reactions.
For biocatalyst preparation we used a commercially available
heavy-duty detergent powder purchased from a local super-
market and used it without any further treatment.
aChongqing Key Laboratory of Catalysis & Functional Organic Molecules, College of
Environment and Resources, Chongqing Technology and Business University,
Chongqing 400067, China
bDepartment of Biotechnology, Del University of Technology, Van der Maasweg 9,
2629HZ Del, The Netherlands. E-mail: F.Hollmann@tudel.nl
† Electronic supplementary information (ESI) available: Details of the
experimental procedures and additional analytic material. See DOI:
10.1039/c9ra05828a
Scheme 1 Selection of typical, preparative applications of lipases in
organic synthesis.
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Fig. 1 ELP driven transesterification of ethyl acetate to isopentyl acetate. (a) The rate dependency of the isopentyl acetate production on
isopentyl alcohol concentration. Conditions: 1000 mg ELP, 60 ꢀC, shaking speed 180 rpm, 24 h. (b) Effect of ELP loading on the trans-
esterification of ethyl acetate. Conditions: c(isopentyl alcohol) ¼ 500 mM, 60 ꢀC, rotating speed 99 rpm, 72 h. (c) Effect of temperature on the
isopentyl acetate formation. Conditions: c(isopentyl alcohol) ¼ 500 mM, 1000 mg ELP, rotating speed 99 rpm, 36 h. (d) Time course of the
isopentyl acetate formation performed by shaking (A) and rotating (>). Conditions: c(isopentyl alcohol) ¼ 500 mM, 1000 mg ELP, 60 ꢀC, shaking
speed 180 rpm or rotating speed 99 rpm. All reactions using ethyl acetate as solvent (ca. 20 eq. on isopentyl alcohol) were performed in
a reaction scale of 10 mL. *: Reactions were performed in an oil bath with stirring speed 300 rpm. Error bars indicate the standard deviation of
duplicate experiments (n ¼ 2).
changing from horizontal to orbital shaking (Fig. S1 and S3†) excellent recyclability. As shown in Fig. 3, the ELP could be
enhanced the reaction rate more than two-fold (Fig. 1d). We recycled at least 8 times maintaining 89.7 ꢁ 1.1% of its initial
therefore assumed that diffusion-limitation may be overall rate- transesterication activity.
limiting. Interestingly enough, decreasing the particle size of
the ELP by mechanical grinding, had no signicant effect on the substrate scope of the ELP-catalysed transesterication and
reaction rate (Fig. S5†). hydrolysis reactions (Table 1). Depending on the steric demand
Encouraged by these results, we further explored the
The preparative applicability of ELP was exemplarily of the starting material, good to poor reaction yields were ob-
demonstrated in the synthesis of isopentyl acetate on litre-scale tained suggesting that ELP may be practical catalysts for at least
(Fig. 2). Over at least 12 days, the product accumulated linearly a range of different esters.
indicating a very high stability of the enzyme under the
We also tried the ELP as catalyst for the kinetic resolution of
comparably harsh reaction conditions. Overall, 1.02 g (isolated racemic 1-phenyl ethanol. Though only one product enantiomer
product) were obtained from this reaction (Fig. S7 and S8†). was observed in the reaction (Fig. S9†), indicating an exclusive
Admittedly, the catalyst performance in this reaction was not enantioselectivity, the very low productivity of the reaction (i.e.
most convincing (150 g ELP per gram of product within 12 the conversion of the secondary alcohol) was discouraging.
days). We therefore investigated the recyclability of the ELP
Finally, we investigated the applicability of the ELP as cata-
under reaction conditions. Very pleasingly, the ELP showed lyst for the epoxidation of styrene. Ever since the rst report by
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Table 1 Substrate scope of ELP driven transesterification, esterifica-
tion and hydrolysis reactionsa
Entry Substrate
Product
c(product)/mM
7.8 ꢁ 0.6
1b
2b
9.2 ꢁ 0.2
Fig. 2 Time course of the preparative-scale synthesis of isopentyl
acetate formation using ELP. Conditions: 1 L reaction scale, c(isopentyl
alcohol) ¼ 500 mM, ethyl acetate as solvent (ca. 20 eq.), 150 g ELP,
60 ꢀC, stirring speed 400 rpm. Error bars indicate the standard devi-
ation of duplicate experiments (n ¼ 2).
3c
0.3
9
¨
Bjorkling et al., chemoenzymatic epoxidation reactions using
4b
4.6 ꢁ 0.4
lipases is enjoying an even increasing popularity.10 Since
modern heavy-duty detergents also contain signicant amounts
of inorganic peroxides we also tested the application of the
chemoenzymatic Prilezhaev reaction on styrene (Table 2).
The chemoenzymatic Prilezhaev reaction necessitates,
next to the lipase catalyst, hydrogen peroxide and a carbox-
ylic acid co-catalysts (being transformed into the corre-
sponding peracid to perform the epoxidation reaction). As
shown in Table 2, ELP apparently contains all these
components as very signicant production of styrene oxide
was observed in the presence of the ELP alone. Adding more
carboxylic acids improved the epoxide yield. It is interesting
5b
0.7
0.7
6b
7d
6.0
2.0
8e
9f
6.2 ꢁ 0.3
a
Reaction conditions: 10 mL reaction scale, 1000 mg ELP, 60 ꢀC,
b
rotating speed 99 rpm, 48 h. c(substrate) ¼ 500 mM, ethyl
c
acetate as solvent. c(substrate) ¼ 100 mM, ethyl acetate as
d
solvent. c(cinnamyl alcohol) ¼ 10 mM, c(vinyl acetate) ¼ 20 mM,
e
toluene as solvent, stirring speed 500 rpm. c(phenylacetic acid)
¼ 10 mM, c(1-heptanol) ¼ 20 mM, toluene as hydrophobic solvent
Fig. 3 Recyclability of ELP for transesterification of ethyl acetate to
isopentyl acetate. Relative activity ¼ c(isopentyl acetate, cycle run) ꢂ
c(isopentyl acetate, first run)ꢃ1. Conditions: 10 mL reaction scale,
c(isopentyl alcohol) ¼ 500 mM, ethyl acetate as solvent (ca. 20 eq.),
1000 mg ELP, 60 ꢀC, rotating speed 99 rpm, 48 h for one cycle. Error
bars indicate the standard deviation of duplicate experiments (n ¼ 2).
f
for phenylacetic acid, stirring speed 500 rpm. 2 mL triolein + 20
mL H2O, 200 mg ELP, stirring speed 500 rpm. Error bars indicate
the standard deviation of duplicate experiments (n ¼ 2).
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Table 2 Epoxidation of styrene by ELPa
nancial possibilities to utilise readily available and cheap
laundry compositions.
Conflicts of interest
There are no conicts to declare.
Acknowledgements
Financial support by Project of China Scholarship Council (No.
201808500035), by the European Research Commission (ERC
consolidator grant, No. 648026), and the Netherlands Organi-
sation for Scientic Research (VICI grant No. 724.014.003) is
gratefully acknowledged.
c(styrene
oxide)/mM
Reactions
Styrene only
3.6
6.2
Styrene + octanoic acidb
a
Notes and references
Reaction conditions: 10 mL styrene solution, c(styrene) ¼ 500 mM,
toluene as solvent, 1000 mg ELP, 60 ꢀC, stirring speed 500 rpm, 48 h.
b
c(octanoic acid) ¼ 100 mM.
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lipase-catalysed formation of the peracid even under non-
aqueous conditions.
4 C. Pusch, Electrophoresis, 1997, 18, 1103–1104.
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Conclusions
With this contribution we aimed at demonstrating the synthetic
applicability of commercially available heavy duty detergent
compositions for the synthesis of esters and epoxides repre-
senting some typical lipase reactions. Modern ELPs contain all
components needed for these reactions and therefore consti-
tute a very cost-efficient alternative to expensive enzyme prep-
arations. We are convinced that further, classical hydrolase
reactions are possible using ELPs.
¨
9 F. Bjorkling, S. E. Godtfredsen and O. Kirk, J. Chem. Soc.,
Chem. Commun., 1990, 19, 1301–1303.
This contribution may inspire school or university teachers 10 B. O. O. Burek, S. Bormann, F. Hollmann, J. Bloh and
and may be an alternative for research groups with limited
D. Holtmann, Green Chem., 2019, 21, 3232–3249.
24042 | RSC Adv., 2019, 9, 24039–24042
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