Table S1 Overview of expression systems for heterologous laccase production in yeasts
Source
classification
Source
Laccase
Genetically engineered
Note to genetically
engineered laccases
Reference
Expression host: Cryptococcus sp. S-2
Fungi, ascomycota
Gaeumannomyces
graminis
LAC2
no
[167]
Fungi,
basidiomycota
Trametes versicolor
LAC5
no
[167]
Coriolopsis gallica
LCC1
no
[162]
Pleurotus ostreatus
POXA3a, POXA3b no
[209]
Pleurotus ostreatus
POXC, POXA1b
no
[122]
Pleurotus ostreatus
POXC, POXA1b
directed evolution by random
mutagenesis
Pleurotus ostreatus
POX3, POX4
no
[123]
Trametes trogii
LCC1
no
[150,210]
LCC1
no
[151,183,204]
Expression host: Kluyveromyces lactis
Fungi,
basidiomycota
higher activity, increased
stability at acidic as well as
alkaline pH
[152,186]
Expression host: Pichia methalonica
Fungi,
basidiomycota
Trametes versicolor
Expression host: Pichia pastoris
Bacteria
Thermus thermophilus
SG0.5JP17-16
LACTt
no
[119]
Fungi, ascomycota
Botrytis aclada
LAC
no
[159]
Monilinia fructigena
LCC2
optimized codon usage
Gaeumannomyces
graminis
LAC2
no
[167]
Yarrowia lipolytica
YlLAC
no
[115]
basidiomycete PM1
strain CECT 2971
PM1
directed evolution by random pH-activity profile shifted to
mutagenesis and/or DNA
the neutral-basic range,
recombination in S. cerevisiae increased resistance to halides
but produced in P. pastoris
[164]
Coprinopsis cinerea
Okayama-7 #130
CcLCC2
no
[158]
Coprinus comatus
LAC1, LAC2
no
[199]
Coprinus comatus
LAC3, LAC4
fusing an additional 10 amino
acids tag at N-terminus
volumetric activities
dramatically enhanced from
undetectable levels
[153]
Coprinus cinereus
LCC5I
optimized codon usage
effective laccase gene
expression
[141]
Cryptococcus
LAC1
no
Fungi,
basidiomycota
effective laccase gene
expression
[200]
[137]
neoformans
Cyathus bulleri
LAC
no
[127]
Cyathus bulleri
LCC
random mutagenesis
Fomes lignosus
LCC
no
Fomes lignosus
LCC
methane sulfonate-based
random mutagenesis in vitro
Flammulina velutipes
LCC
no
[202]
Ganoderma fornicatum
0814
LAC1
no
[131]
Ganoderma lucidum
GlLCC1
optimized codon usage, GC
effective constitutive gene
content and regions generating expression under GAP
secondary structures
promoter and α-factor leader
in P. pastoris
[207]
Ganoderma lucidum
LAC1
no
[166]
Ganoderma lucidum
7071-9
LCCI
optimized codon usage
Ganoderma sp. En3
LAC-En3-1
no
[142,156]
Lentinus sp.
LCC4
no
[109]
increased ability to decolorize [155]
reactive blue 21 (RB21), a
phthalocyanine dye reported to
be decolorized only by
peroxidases
[110]
improved expression and kcat
effective laccase gene
expression
[206]
[134]
Lentinus sp.
LCC4
site-directed mutagenesis
possibility to evaluate the
effect on the laccase activity
by single glycosylation site
deficiency
[109]
Lenzites gibbosa
LAC
no
Physisporinus rivulosus
LAC1, LAC2
site-directed mutagenesis
Pleuotus ostreatus
LCCPol
no
[135]
Pleuotus ostreatus
POXA1B
optimized codon usage, GC
effective constitutive gene
content and regions generating expression under GAP
secondary structures
promoter and α-factor leader
in P. pastoris
[207]
Pleurotus sajor-caju
P32-1
LAC4
no
[133]
Polyporus
grammocephalus TR16
LAC
no
[130]
Pycnoporus sanguineus
LAC
no
[145]
Pycnoporus sanguineus
H275
LCC1
no
[161]
Pycnoporus cinnabarinus LAC1
I-937
no
[201]
Trametes sanguinea
M85-2
LCC1-5
no
[129]
Trametes sp. AH28-2
LACB
no
[148]
[144]
increased specific laccase
activity
[128]
Trametes sp. AH28-2
LACB
mutagenesis by low-energy
nitrogen ion implantation
increased specific laccase
activity, light change in its
catalytic ability but superior
thermal stability
Trametes sp. 420
LACC
no
[146]
Trametes sp. 420
LACD
no
[126,147]
Trametes sp. 420
LACD
site-directed mutagenesis
Trametes sp. 48424
LAC48424-1
no
[143]
Trametes sp. 5930
LAC5930-1
no
[138,140]
Trametes trogii
LCC1, LCC2
no
[28,226,227]
Trametes versicolor
LCC1, LCC2
no
[120,170,208,
223]
Trametes versicolor
LCCI
site-directed mutagenesis
Trametes versicolor
LCCA, LCCB,
LCCC
no
[154,157,203]
Trametes versicolor
LCCIV
no
[125]
mutants with different laccase
activity
redox potential of the active
site shifted to more negative
values, enhanced rate of
electron transfer between an
oxidoreductase and an
electrode
[169]
[163]
[216]
Chromista,
oomycota
Plants
Trametes versicolor
LAC5
no
[167]
Phythophthora capsici
LAC1
no
[118]
Phythophthora capsici
LAC2
no
[117]
Gossypium arboreum
LAC1
no
[43]
Cryphonectria parasitica LAC3
no
[192]
Melanocarpus albomyces LAC1
site-directed mutagenesis
confirmation of the critical
role of the last amino acids in
the C-terminus of MaLAC1
Myceliophthora
thermophila
MtL
directed evolution by random
mutagenesis
improved expression,
[30,194]
increased kcat, enhanced total
activity, substrate specificity
not changed, higher activity in
the presence of organic
solvents
Myceliophthora
thermophila
MtL
combinatorial saturation
mutagenesis through in vivo
overlap extension (IVOE)
higher kinetic parameters,
resistance to high
concentrations of organic
solvents
Expression host: Saccharomyces cerevisiae
Fungi, ascomycota
[84]
[113,215]
Fungi,
basidiomycota
Myceliophthora
thermophila
MtL
in vivo recombine mutant
libraries with different
nucleotide bias created by in
vitro evolution under the
selective pressure of
increasing concentrations of
organic cosolvents
higher kinetic parameters,
[195]
resistance to high
concentrations of organic
solvents, the electron transfer
pathway between the reducing
substrate and the T1 copper
ion altered, improved catalytic
efficiency towards nonphenolic and phenolic
substrates, redox potential not
significantly altered
basidiomycete PM1
strain CECT 2971
PM1
directed evolution by random
mutagenesis and/or DNA
recombination
enhanced total activity,
enhanced functional
expression, highly active and
stable enzyme in terms of
temperature, pH range, and
organic cosolvents, improved
catalytic capacities
Coprinus congregatus
LAC2
no
[139]
Coriolus hirsutus
PO1, PO2
no
[193]
Lentinula edodes
LCC1, LCC4
no
[189]
Pleurotus eryngii
ERY3
no
[188]
Pleurotus eryngii
ERY4
site-directed mutagenesis
analysis the role of the Cterminus of ERY4 protein
[112]
[124]
Pleurotus eryngii
ERY3/ERY4
(4NC3)
DNA shuffling by DNA
recombination to form
chimeric laccases from
different laccase isoforms,
displayed on the cell surface
of S. cerevisiae
chimeras with best
performances in terms of
enzymatic activities, affinities
for different substrates and
stability at a broad range of
temperatures and pHs, S.
cerevisiae surface display
[222]
Pleurotus ostreatus
POXC,POXA1b
no
Pleurotus ostreatus
POXC,POXA1b
directed evolution by random
mutagenesis
higher activity, increased
stability at acidic as well as
alkaline pH
[152,186]
Pleurotus ostreatus
POXA1b
site-directed mutagenesis
analysis the role of the Cterminal tail of POXA1b in
affecting its catalytic and
stability properties
[29]
Pleurotus ostreatus
POX3, POX4
no
[122]
[123]
Pycnoporus cinnabarinus LAC1
directed evolution by random
mutagenesis combined with
site-directed mutagenesis
and/or in vivo overlap
extension (IVOE)
enhanced total activity,
[168]
improved secretion levels due
to the evolved α-factor,
increased kcat, pH activity
profile shifted to more neutral
values, the thermostability and
the broad substrate specificity
retained
Pycnoporus
cinnabarinus/PM1
basidiomycete
DNA shuffling by DNA
recombination to form
chimeric laccases from two
different fungi
active hybrid laccases with
combined characteristics in
terms of pH activity and
thermostability
PcLAC1/PM1
[187]
Plants
Pycnoporus coccineus
LCC1
no
[196]
Trametes sp. AH28-2
LACA
no
[198]
Trametes sp. C30
LAC1-5
no
[171,191,197]
Trametes sp. C30
LAC3
optimized codon usage
effective laccase gene
expression
Trametes sp. C30
LAC3/LAC1,
LAC3/LAC2,
LAC3/LAC5
DNA shuffling by DNA
recombination to form
chimeric laccases from
different laccase isoforms
pH tolerance extended towards [31]
alkaline pH values, increased
kcat
Trametes sp. Ha1
laccase I
site-directed mutagenesis,
displayed on the cell surface
of S. cerevisiae
S. cerevisiae surface display
Trametes versicolor
LCC1, LCC2
no
[121,160]
Trametes versicolor
LCCIII
no
[136]
Trametes versicolor
AUMH8272
LCCα
no
[132]
Trametes sanguinea
M85-2
LCC1
no
[129]
Pinus taeda
LAC2
no
[116]
Pycnoporus cinnabarinus LAC1
no
[212]
[190]
[225]
Expression host: Yarrowia lipolytica
Fungi,
basidiomycota
Trametes versicolor
IIIb
no
[213]
Trametes versicolor
IIIb
site-directed mutagenesis
increased kcat, optimal pH
shifted
[214]
Trametes versicolor
DSM11269
LCC1
directed evolution combining
random and site-directed
mutagenesis in Y.lipolytica
enhanced secreted enzyme
activity, increased catalytic
efficiency
[149]