1
Supplementary information
H. jecorina
cellobiohydrolase I
stabilizing mutations
identified using
non
-
contiguous recombination.
Matthew A. Smith,
Claire
N.
Bedbrook, Timothy Wu
, Frances H. Arnold
Division of Chemistry and Chemical Engineering,
California Institute of Technology,
Pasadena, CA, 91125, USA.
2
Supplementary Figure 1
.
Thermostabilities of a maximally informative subset
(32
chimeras)
of the library.
The
T
R50
is t
he incubation temperature at which a chimera's
residual activity
after 10 minutes
is half its
activity before incubation
. Measurements
were performed in duplicate. The parental enzymes are highlighted.
45
50
55
60
65
Thermostability, T
R50
(°C)
Chimeras
P1
P2
P3
3
Supplementary Figure 2
T
R50
measurements for a range of DTT concentrations
.
A)
P
arental cellulase
T. emersonii
CBH
I
recovers
some
activity
after incubation if there
is no DTT present. The enzyme does not
recover activity
when incubated with
1
mM
DTT. B)
C
himeric cellulase
222211222322
is able to
recover some activity
after
incubation
with
1
mM DTT.
A
B
T.
emersonii
CBH1
Chimera 222211222322
4
Supplementary Table 1
Amino acid
sequences of the
maximally informative subset
of 35
chimeric cellulases
.
T
he
H. jecorina
CBH
I
linker and cellulose binding domain
are attached to the C
-
terminus
. The chimera nomenclature is a series of numbers, each
represe
nting a parent for the blocks A
-
L. For example, chimera 123113322331 has
parent 1's sequence for block A, parent 2 for block B, etc.
The three chimeras with no
detectable activity are highlighted w
ith an asterisk
.
123113322331
QQACTLQSETHPPLTWKRCSSGGTCSTVNGSVTIDANWRWTHTVSGSTNCYDGNTWDTSLCTDDKSCAQ
TCCLDGADYSSTYGITTSGDSLNLKFVTQSAQKNVGSRVYLMASDTTYQMFELLNREFTFDVDVSNLPC
GLNGALYFVSMDADGGMSKYSGNKAGAKYGTGYCDAQCPRDLKFIDGEANVEGWQPSSNNANTGIGDHG
SCCSEMDVWEANSISEAVTPHPCTTVGQTMCSGDDCGGTYSDNRYGGVC
DPDGCDFNPYRMGNTSFYGP
GMTVDTTKKMTVVTQFLTDDGTDTGTLSEIKRFYVQDGKIIAQPNSDISGVTGNSITTEFCTAQKQAFS
NTDDFNRKGGLAQMSKALAGPMVLVMSVWDDYYAQMLWLDSTYPTNETSSTPGAVRGSCPTDSGVPAQV
ESQSPNSNVIFSNIRFGPIGSTVPGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCA
SGTTCQVLNPYYSQCL
21121323311
2
QSACTLQSETHPPLTWQKCTAPGSCTQQTGSVTIDANWRWVHATNSSTNCYTGNTWSSTLCPDNETCAK
NCCLDGADYSSTYGITTSGNSLNLKFVTGSNVGSRVYLMASDTTYQIFKLLGNEFSFDVDVSNLPCGLN
GALYFVAMDADGGVSKYPTNTAGAKYGTGYCDSQCPRDLKFINGQANVEGWEPSSNNANTGIGGHGSCC
AEMDVWEANSISNAFTPHPCDTPGQEICEGDGCGGTYSDNRYGGTC
DPDGCDFNPYRQGNTSFYGPGSS
FTLDTTKKLTVVTQFHKNSAGVLSEIKRYYVQNGVTFQQPNSKIPGNPGNSITQEYCDAQEVAFGGSSF
SDHGGMAKMGAAMQQGMVLVMSVWDDYAANMLWLDSDYPTDADPTTPGIARGTCSTSSGVPSDVESQSP
NSNVIFSNIKFGPIGSTGNPSGGNPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCAS
GTTCQVLNPYYSQCL
232232332
132
QQACSLTAENHPSLTWQKCTAPGSCTQQTGAVVLDANWRWVHDVNGYTNCYTGNTWDPTYCPDGETCAQ
NCCLDGADYEGTYGVTTSGNSLKLNFVTGSNVGSRLYLMENDTKYQMFELLGNEFSFDVDVSNLPCGLN
GALYFVSMDADGGVSKYPNNKAGAKYGTGYCDAQCPRDLKFINGEANVGNWTPSTNNANTGIGRYGSCC
SEMDVWEANSISNAVTPHPCTTVGQSRCEADTCGGTYSSDRYAG
VCDPDGCDFNPYRMGNTSFYGPGMT
VDTTKKMTVVTQFHKNSAGVLSEIKRFYVQDGKIIAQPNSDISGVTGNSITTEFCTAQEQAFGGSSFSD
KGGLAQMSKALAGPMVLVMSLWDDYAANMLWLDSDYPTDADPTTPGIARGTCPTTSGVPSDVESQSPNS
YVTYSNIKFGPIGSTVPPSGGNPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGT
TCQVLNPYYSQCL
322321222
212
QQACTATAENHPPLTWQECTSGGSCTTQNGAVVIDANWRWTHDVNGYTNCYTGNQWDPTYCPDDETCAQ
NCCVDGAAYASTYGVTSSGSSLSIGFVTKHQYGTNVGARLYLLQDDSTYQIFKLLNREFSFDVDVSQLP
CGLNGALYFVAMDADGGVSKYPNNKAGAKYGTGYCDSQCPRDLKFIDGEANVEGWQPSSNDANAGFGDH
GSCCAEMDVWEANSISNAVTPHPCDTPGQTMCSGDDCGGTYSND
RYAGTCDPDGCDFNPYRMGNTSFYG
PGSSFTLDTTKKLTVVTQFLTDDGTDTGTLSEIKRYYVQNGVTFQQPNSDISGVTGNSITTEFCTAQKQ
AFGDTDDFSQHGGLAKMGAAMQQGMVLVMSLWDDHYAQMLWLDSTYPIDQAGAPGAERGTCPTDSGVPA
EIEAQSPNAKVTFSNIKFGPIGSTGNASPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPT
VCASGTTCQVLNPYYSQCL
5
33
1212211233 *
QSACTLQSETHPPLTWQECTAPGSCTTQNGSVVLDANWRWVHATNSSTNCYTGNQWSSTLCPDGETCAK
NCCLDGADYEGTYGVTTSGSSLKLNFVTGSNVGSRLYLMASDTTYQIFKLLGNEFSFDVDVSNLGCGLN
GALYFVAMDADGGVSKYPTNTAGAKYGTGYCDSQCPRDLKFINGEANVGNWTPSTNDANAGFGRYGSCC
AEMDIWEANNMATALTPHPCDTPGQSRCEADTCGG
TYSDNRYGGTCDPDGCDWNAYRLGDKTFYGKGMT
VDTNKKMTVVTQFETSGAINRFYVQDGKIIANAEAELGSYSGNELNDDYCTAEKAEFGDTDDFSQHGGL
AKMGAAMQQGMVLVMSLWDDHYANMLWLDSTYPIDQAGAPGAERGACPTTSGVPSDVESQVPNSYVTYS
NIKFGPIGSTVPASPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGTTCQVLNPY
YSQCL
311112222333
QSACTLQSETHPPLTWKRCSSGGTCSTVNGSVVLDANWRWTHATNSSTNCYTGNQWSSTLCPDNETCAK
NCCLDGADYEGTYGVTTSGDSLKLNFVTQSAQKNVGSRLYLMASDTTYQIFKLLNREFTFDVDVSNLGC
GLNGALYFVAMDADGGVSKYPTNTAGAKYGTGYCDSQCPRDLKFINGQANVEGWEPSSNDANAGFGGHG
SCCAEMDVWEANNMATAVTPHPCDTPGQEICEGDG
CGGTYSDNRYGGTCDPDGCDFNAYRMGDKTFYGK
GMTVDTNKKMTVVTQFLTDDGTDTGTLSEIKRFYVQDGKIIANAESDISGVTGNSITTEFCTAQKQAFS
NTDDFNRHGGLAKMGAAMQQGMVLVMSLWDDHYANMLWLDSTYPIDQAGAPGAERGACSTSSGVPAQVE
SQVPNSYVTYSNIRFGPIGSTVPGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCAS
GTTCQVLNPYYSQCL
331211323222
QSACTLQSETHPPLTWQECTAPGSCTTQNGSVVIDANWRWVHATNSSTNCYTGNQWSSTLCPDGETCAK
NCCLDGAAYASTYGVTTSGSSLSIGFVTGSNVGARLYLMASDTTYQMFELLNREFSFDVDVSQLPCGLN
GALYFVSMDADGGVSKYPTNTAGAKYGTGYCDAQCPRDLKFINGEANVGNWTPSTNDANAGFGRYGSCC
SEMDVWEANSISNAFTPHPCTTVGQSRCEADTCGGTYSDNRYGGVCDPD
GCDFNPYRQGNTSFYGPGKI
IDTTKPFTVVTQFLTDDGTDTGTLSEIKRFYIQNSNVIPQPNSKIPGNPGNSITQEYCDAQKVAFGDTD
DFSQKGGMAQMSKALAGPMVLVMSLWDDHYANMLWLDSTYPIDQAGAPGAERGTCPTTSGVPSDVESQS
PNAKVTFSNIKFGPINSTFTASPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGT
TCQVLNPYYSQCL
211331132321
QSACSLTAENHPSLTWKRCTSGGSCSTVNGAVVIDANWRWTHATNSSTNCYTGNTWSSTLCPDNETCAK
NCCVDGAAYASTYGVTTSGDSLSIGFVTKHQYGTNVGARLYLMENDTKYQEFTLLGNEFTFDVDVSQLP
CGLNGALYFVSMDADGGVSKYPTNTAGAKYGTGYCDSQCPRDLKFINGQANVEGWEPSSNNANTGIGGH
GSCCSEMDVWEANSISEAVTPHPCTTVGQEICEGDGCGGTYSSDRYAGT
CDPDGCDFNPYRMGNTSFYG
PGKIIDTTKPFTVVTQFHKNSAGVLSEIKRFYIQNSNVIPQPNSDISGVTGNSITTEFCTAQKQAFSNT
DDFNRKGGLTQFKKATSGGMVLVMSLWDDYAANMLWLDSDYPTDADPTTPGIARGSCSTSSGVPAEIEA
QSPNAKVTFSNIRFGPINSTFTGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASG
TTCQVLNPYYSQCL
212322323311
QQACTATAENHPPLTWKRCTSGGSCSTVNGAVVLDANWRWTHDVNGYTNCYTGNTWDPTYCPDNETCAQ
NCCVDGADYEGTYGVTSSGDSLKLNFVTKHQYGTNVGSRLYLLQDDSTYQMFELLNREFTFDVDVSNLP
CGLNGALYFVSMDADGGVSKYPNNKAGAKYGTGYCDAQCPRDLKFINGQANVEGWEPSSNNANTGIGGH
GSCCSEMDVWEANSISEAFTPHPCTTVGQEICEGDGCGGTYSNDRYAGV
CDPDGCDFNPYRQGNTSFYG
PGSSFTLDTTKKLTVVTQFLTDDGTDTGTLSEIKRYYVQNGVTFQQPNSKIPGNPGNSITQEYCDAQKV
AFSNTDDFNRKGGMAQMSKALAGPMVLVMSLWDDYAANMLWLDSDYPTDADPTTPGIARGSCSTSSGVP
AEIEAQSPNSYVTYSNIRFGPIGSTGNGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPT
VCASGTTCQVLNPYYSQCL
6
32321211
1312
QQACTLQSETHPPLTWKRCTAPGSCSTVNGSVVLDANWRWVHTVSGSTNCYTGNQWDTSLCTDDKSCAQ
TCCLDGADYEGTYGVTTSGDSLKLNFVTGSNVGSRLYLMASDTTYQEFTLLGNEFTFDVDVSNLPCGLN
GALYFVSMDADGGMSKYSGNKAGAKYGTGYCDSQCPRDLKFIDGEANVEGWQPSSNDANAGFGDHGSCC
SEMDIWEANSISNALTPHPCTTVGQTMCSGDDCGGTYSDNRYG
GTCDPDGCDWNPYRLGNTSFYGPGSS
FTLDTTKKLTVVTQFETSGAINRYYVQNGVTFQQPNAELGSYSGNELNDDYCTAEKAEFSNTDDFNRKG
GLTQFKKATSGGMVLVMSLWDDHYAQMLWLDSTYPIDQAGAPGAERGTCPTDSGVPSDVESQSPNSYVT
YSNIRFGPIGSTGNGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGTTCQVLNP
YYSQCL
312223311121
QQ
ACTATAENHPPLTWQKCTAPGSCTQQTGAVTIDANWRWVHDVNGYTNCYTGNQWDPTYCPDNETCAQ
NCCLDGADYSSTYGITSSGNSLNLKFVTGSNVGSRVYLLQDDSTYQMFELLGNEFSFDVDVSNLPCGLN
GALYFVSMDADGGVSKYPNNKAGAKYGTGYCDAQCPRDLKFINGQANVEGWEPSSNDANAGFGGHGSCC
SEMDIWEANSISEALTPHPCTTVGQEICEGDGCGGTYSNDRYAGVCDPDGC
DWNPYRLGNTSFYGPGKI
IDTTKPFTVVTQFETSGAINRFYIQNSNVIPQPNAELGSYSGNELNDDYCTAEEAEFGGSSFSDKGGLA
QMSKALAGPMVLVMSVWDDHYANMLWLDSTYPIDQAGAPGAERGSCSTSSGVPSDVESQSPNSNVIFSN
IKFGPINSTFTPSGGNPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGTTCQVLN
PYYSQCL
221221333333
QSACTATAE
NHPPLTWKRCTAPGSCSTVNGAVVIDANWRWVHATNSSTNCYTGNTWSSTLCPDDETCAK
NCCLDGAAYASTYGVTSSGDSLSIGFVTGSNVGARLYLLQDDSTYQMFELLGNEFTFDVDVSQLGCGLN
GALYFVSMDADGGVSKYPTNTAGAKYGTGYCDAQCPRDLKFIDGEANVEGWQPSSNNANTGIGDHGSCC
SEMDVWEANNMATAFTPHPCTTVGQTMCSGDDCGGTYSNDRYAGVCDPDGCDFNAYRQ
GDKTFYGKGMT
VDTNKKMTVVTQFHKNSAGVLSEIKRFYVQDGKIIANAESKIPGNPGNSITQEYCDAQKVAFSNTDDFN
RKGGMAQMSKALAGPMVLVMSLWDDYAAQMLWLDSDYPTDADPTTPGIARGACPTDSGVPSDVESQVPN
AKVTFSNIRFGPIGSTVPGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGTTCQ
VLNPYYSQCL
322132333121
QQACSLTAENHPS
LTWQKCSSGGTCTQQTGAVVLDANWRWTHDVNGYTNCYTGNQWDPTYCPDDETCAQ
NCCLDGADYEGTYGVTTSGNSLKLNFVTQSAQKNVGSRLYLMENDTKYQMFELLGNEFSFDVDVSNLPC
GLNGALYFVSMDADGGVSKYPNNKAGAKYGTGYCDAQCPRDLKFIDGEANVEGWQPSSNDANAGFGDHG
SCCSEMDVWEANSISEAFTPHPCTTVGQTMCSGDDCGGTYSSDRYAGVCDPDGCDFNPYRQG
NTSFYGP
GKIIDTTKPFTVVTQFHKNSAGVLSEIKRFYIQNSNVIPQPNSKIPGNPGNSITQEYCDAQEVAFGGSS
FSDKGGMAQMSKALAGPMVLVMSLWDDHYAQMLWLDSTYPIDQAGAPGAERGSCPTDSGVPAQVESQSP
NSYVTYSNIKFGPINSTFTPSGGNPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCAS
GTTCQVLNPYYSQCL
231123332212
QSACTATAENHP
PLTWQECSSGGTCTTQNGAVTIDANWRWTHATNSSTNCYTGNTWSSTLCPDGETCAK
NCCLDGADYSSTYGITSSGSSLNLKFVTQSAQKNVGSRVYLLQDDSTYQMFELLGNEFSFDVDVSNLPC
GLNGALYFVSMDADGGVSKYPTNTAGAKYGTGYCDAQCPRDLKFINGEANVGNWTPSTNNANTGIGRYG
SCCSEMDVWEANSISNAVTPHPCTTVGQSRCEADTCGGTYSNDRYAGVCDPDGCDFNPYRM
GNTSFYGP
GSSFTLDTTKKLTVVTQFHKNSAGVLSEIKRYYVQNGVTFQQPNSDISGVTGNSITTEFCTAQKQAFGD
TDDFSQKGGLAQMSKALAGPMVLVMSVWDDYAANMLWLDSDYPTDADPTTPGIARGTCPTTSGVPAQVE
SQSPNSNVIFSNIKFGPIGSTGNASPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCA
SGTTCQVLNPYYSQCL
7
121233122211
QSACSLTAENHPSLTWQECTAPGSCTTQNGAVTIDANWRWVHATNSSTNCYDGNTWSSTLCPDDETCAK
NCCLDGADYSSTYGITTSGSSLNLKFVTGSNVGSRVYLMENDTKYQEFTLLNREFSFDVDVSNLPCGLN
GALYFVSMDADGGVSKYPTNTAGAKYGTGYCDSQCPRDLKFIDGEANVEGWQPSSNNANTGIGDHGSCC
SEMDVWEANSISEAVTPHPCTTVGQTMCSGDDCGGTYSSDRYAGTCDPD
GCDFNPYRMGNTSFYGPGSS
FTLDTTKKLTVVTQFLTDDGTDTGTLSEIKRYYVQNGVTFQQPNSDISGVTGNSITTEFCTAQKQAFGD
TDDFSQKGGLTQFKKATSGGMVLVMSVWDDYYAQMLWLDSTYPTNETSSTPGAVRGSCPTDSGVPSDVE
SQSPNSNVIFSNIKFGPIGSTGNASPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCA
SGTTCQVLNPYYSQCL
221332311112
QSACSLTAENHPSLTWQKCTSGGSCTQQTGAVVLDANWRWTHATNSSTNCYTGNTWSSTLCPDDETCAK
NCCVDGADYEGTYGVTTSGNSLKLNFVTKHQYGTNVGSRLYLMENDTKYQMFELLGNEFSFDVDVSNLP
CGLNGALYFVSMDADGGVSKYPTNTAGAKYGTGYCDAQCPRDLKFIDGEANVEGWQPSSNNANTGIGDH
GSCCSEMDIWEANSISNALTPHPCTTVGQTMCSGDDCGGTYSSDRYA
GVCDPDGCDWNPYRLGNTSFYG
PGSSFTLDTTKKLTVVTQFETSGAINRYYVQNGVTFQQPNAELGSYSGNELNDDYCTAEEAEFGGSSFS
DKGGLAQMSKALAGPMVLVMSLWDDYAAQMLWLDSDYPTDADPTTPGIARGTCPTDSGVPAEIEAQSPN
SYVTYSNIKFGPIGSTGNPSGGNPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASG
TTCQVLNPYYSQCL
212312332233
QQACTLQSETHPPLTWQECTSGGSCTTQNGSVVLDANWRWTHDVNGYTNCYTGNTWDPTYCPDNETCAQ
NCCVDGADYEGTYGVTTSGSSLKLNFVTKHQYGTNVGSRLYLMASDTTYQMFELLGNEFSFDVDVSNLG
CGLNGALYFVSMDADGGVSKYPNNKAGAKYGTGYCDAQCPRDLKFINGQANVEGWEPSSNNANTGIGGH
GSCCSEMDVWEANNMATAVTPHPCTTVGQEICEGDGCGGTYSDNRYG
GVCDPDGCDFNAYRMGDKTFYG
KGMTVDTNKKMTVVTQFHKNSAGVLSEIKRFYVQDGKIIANAESDISGVTGNSITTEFCTAQKQAFGDT
DDFSQKGGLAQMSKALAGPMVLVMSLWDDYAANMLWLDSDYPTDADPTTPGIARGACSTSSGVPAEIEA
QVPNSYVTYSNIKFGPIGSTVPASPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCAS
GTTCQVLNPYYSQCL
31332233212
2
QQACTATAENHPPLTWQKCTSGGSCTQQTGAVVLDANWRWTHTVSGSTNCYTGNQWDTSLCTDNKSCAQ
TCCVDGADYEGTYGVTSSGNSLKLNFVTKHQYGTNVGSRLYLLQDDSTYQMFELLGNEFSFDVDVSNLP
CGLNGALYFVSMDADGGMSKYSGNKAGAKYGTGYCDAQCPRDLKFINGQANVEGWEPSSNDANAGFGGH
GSCCSEMDVWEANSISNAVTPHPCTTVGQEICEGDGCGGTYSNDRY
AGVCDPDGCDFNPYRMGNTSFYG
PGKIIDTTKPFTVVTQFHKNSAGVLSEIKRFYIQNSNVIPQPNSDISGVTGNSITTEFCTAQEQAFGGS
SFSDKGGLAQMSKALAGPMVLVMSLWDDHYANMLWLDSTYPIDQAGAPGAERGTCSTSSGVPAEIEAQS
PNSYVTYSNIKFGPINSTFTPSGGNPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCA
SGTTCQVLNPYYSQCL
213131232
211
QQACSLTAENHPSLTWQECSSGGTCTTQNGAVVIDANWRWTHTVSGSTNCYTGNTWDTSLCTDNKSCAQ
TCCLDGAAYASTYGVTTSGSSLSIGFVTQSAQKNVGARLYLMENDTKYQIFKLLGNEFSFDVDVSQLPC
GLNGALYFVAMDADGGMSKYSGNKAGAKYGTGYCDSQCPRDLKFINGQANVEGWEPSSNNANTGIGGHG
SCCAEMDVWEANSISEAVTPHPCDTPGQEICEGDGCGGTYSSDR
YAGTCDPDGCDFNPYRMGNTSFYGP
GSSFTLDTTKKLTVVTQFHKNSAGVLSEIKRYYVQNGVTFQQPNSDISGVTGNSITTEFCTAQKQAFGD
TDDFSQHGGLAKMGAAMQQGMVLVMSLWDDYAANMLWLDSDYPTDADPTTPGIARGSCSTSSGVPAQVE
SQSPNAKVTFSNIKFGPIGSTGNASPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCA
SGTTCQVLNPYYSQCL
8
131232
232321
QSACSLTAENHPSLTWKRCTAPGSCSTVNGAVVLDANWRWVHATNSSTNCYDGNTWSSTLCPDGETCAK
NCCLDGADYEGTYGVTTSGDSLKLNFVTGSNVGSRLYLMENDTKYQIFKLLGNEFTFDVDVSNLPCGLN
GALYFVAMDADGGVSKYPTNTAGAKYGTGYCDSQCPRDLKFINGEANVGNWTPSTNNANTGIGRYGSCC
AEMDVWEANSISEAVTPHPCDTPGQSRCEADTCGGTYSSDR
YAGTCDPDGCDFNPYRMGNTSFYGPGKI
IDTTKPFTVVTQFHKNSAGVLSEIKRFYIQNSNVIPQPNSDISGVTGNSITTEFCTAQKQAFSNTDDFN
RHGGLAKMGAAMQQGMVLVMSLWDDYYANMLWLDSTYPTNETSSTPGAVRGSCPTTSGVPSDVESQSPN
SYVTYSNIRFGPINSTFTGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGTTCQ
VLNPYYSQCL
1121313223
22
QQACSLTAENHPSLTWKRCSSGGTCSTVNGAVVIDANWRWTHDVNGYTNCYDGNTWDPTYCPDNETCAQ
NCCLDGAAYASTYGVTTSGDSLSIGFVTQSAQKNVGARLYLMENDTKYQMFELLNREFTFDVDVSQLPC
GLNGALYFVSMDADGGVSKYPNNKAGAKYGTGYCDAQCPRDLKFINGQANVEGWEPSSNNANTGIGGHG
SCCSEMDVWEANSISNAVTPHPCTTVGQEICEGDGCGGTYSSDRY
AGVCDPDGCDFNPYRMGNTSFYGP
GKIIDTTKPFTVVTQFLTDDGTDTGTLSEIKRFYIQNSNVIPQPNSDISGVTGNSITTEFCTAQKQAFS
NTDDFNRKGGLAQMSKALAGPMVLVMSLWDDYYANMLWLDSTYPTNETSSTPGAVRGTCSTSSGVPAQV
ESQSPNAKVTFSNIRFGPINSTFTGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCA
SGTTCQVLNPYYSQCL
11112233
2232
QSACTATAENHPPLTWQECSSGGTCTTQNGAVVLDANWRWTHATNSSTNCYDGNTWSSTLCPDNETCAK
NCCLDGADYEGTYGVTSSGSSLKLNFVTQSAQKNVGSRLYLLQDDSTYQMFELLGNEFSFDVDVSNLPC
GLNGALYFVSMDADGGVSKYPTNTAGAKYGTGYCDAQCPRDLKFINGQANVEGWEPSSNNANTGIGGHG
SCCSEMDVWEANSISNAVTPHPCTTVGQEICEGDGCGGTYSND
RYAGVCDPDGCDFNPYRMGNTSFYGP
GMTVDTTKKMTVVTQFHKNSAGVLSEIKRFYVQDGKIIAQPNSDISGVTGNSITTEFCTAQKQAFGDTD
DFSQKGGLAQMSKALAGPMVLVMSLWDDYYANMLWLDSTYPTNETSSTPGAVRGTCSTSSGVPAQVESQ
SPNSYVTYSNIKFGPIGSTVPASPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASG
TTCQVLNPYYSQCL
31121133
1111
QSACTLQSETHPPLTWQKCTAPGSCTQQTGSVVIDANWRWVHATNSSTNCYTGNQWSSTLCPDNETCAK
NCCLDGAAYASTYGVTTSGNSLSIGFVTGSNVGARLYLMASDTTYQMFELLGNEFSFDVDVSQLPCGLN
GALYFVSMDADGGVSKYPTNTAGAKYGTGYCDAQCPRDLKFINGQANVEGWEPSSNDANAGFGGHGSCC
SEMDIWEANSISEALTPHPCTTVGQEICEGDGCGGTYSDNRYG
GVCDPDGCDWNPYRLGNTSFYGPGSS
FTLDTTKKLTVVTQFHKNSAGVLSEIKRYYVQNGVTFQQPNAELGSYSGNSLNDDYCTAEEAEFGGSSF
SDKGGLAQMSKALAGPMVLVMSLWDDHYANMLWLDSTYPIDQAGAPGAERGSCSTSSGVPSDVESQSPN
AKVTFSNIKFGPIGSTGNPSGGNPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASG
TTCQVLNPYYSQCL
12212213
3333 *
QQACTATAENHPPLTWKRCSSGGTCSTVNGAVVLDANWRWTHDVNGYTNCYDGNTWDPTYCPDDETCAQ
NCCLDGADYEGTYGVTSSGDSLKLNFVTQSAQKNVGSRLYLLQDDSTYQEFTLLGNEFTFDVDVSNLGC
GLNGALYFVSMDADGGVSKYPNNKAGAKYGTGYCDSQCPRDLKFIDGEANVEGWQPSSNNANTGIGDHG
SCCSEMDVWEANNMATAFTPHPCTTVGQTMCSGDDCGGTYS
NDRYAGTCDPDGCDFNAYRQGDKTFYGK
GMTVDTNKKMTVVTQFHKNSAGVLSEIKRFYVQDGKIIANAESKIPGNPGNSITQEYCDAQKVAFSNTD
DFNRKGGMTQFKKATSGGMVLVMSLWDDYYAQMLWLDSTYPTNETSSTPGAVRGACPTDSGVPAQVESQ
VPNSYVTYSNIRFGPIGSTVPGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGT
TCQVLNPYYSQCL
9
233122223223
QQACTATAENHPPLTWQECSSGGTCTTQNGAVVLDANWRWTHTVSGSTNCYTGNTWDTSLCTDGKSCAQ
TCCLDGADYEGTYGVTSSGSSLKLNFVTQSAQKNVGSRLYLLQDDSTYQIFKLLNREFSFDVDVSNLGC
GLNGALYFVAMDADGGMSKYSGNKAGAKYGTGYCDSQCPRDLKFINGEANVGNWTPSTNNANTGIGRYG
SCCAEMDVWEANNMATAFTPHPCDTPGQSRCEADT
CGGTYSNDRYAGTCDPDGCDFNAYRQGDKTFYGK
GKIIDTNKPFTVVTQFLTDDGTDTGTLSEIKRFYIQNSNVIPNAESKIPGNPGNSITQEYCDAQKVAFG
DTDDFSQHGGMAKMGAAMQQGMVLVMSLWDDYAANMLWLDSDYPTDADPTTPGIARGACPTTSGVPAQV
ESQVPNSYVTYSNIKFGPINSTFTASPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVC
ASGTTCQVLNPYYSQ
CL
333222211111
QQACTATAENHPPLTWQKCTAPGSCTQQTGAVVLDANWRWVHTVSGSTNCYTGNQWDTSLCTDGKSCAQ
TCCLDGADYEGTYGVTSSGNSLKLNFVTGSNVGSRLYLLQDDSTYQIFKLLGNEFSFDVDVSNLPCGLN
GALYFVAMDADGGMSKYSGNKAGAKYGTGYCDSQCPRDLKFINGEANVGNWTPSTNDANAGFGRYGSCC
AEMDIWEANSISEALTPHPCDTPGQSRCEADTCGGTYSNDRYAGTCDPD
GCDWNPYRLGNTSFYGPGSS
FTLDTTKKLTVVTQFETSGAINRYYVQNGVTFQQPNAELGSYSGNELNDDYCTAEEAEFGGSSFSDHGG
LAKMGAAMQQGMVLVMSLWDDHYANMLWLDSTYPIDQAGAPGAERGSCPTTSGVPSDVESQSPNSYVTY
SNIKFGPIGSTGNPSGGNPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGTTCQV
LNPYYSQCL
231333111132
QSACS
LTAENHPSLTWQKCTSGGSCTQQTGAVTIDANWRWTHATNSSTNCYTGNTWSSTLCPDGETCAK
NCCVDGADYSSTYGITTSGNSLNLKFVTKHQYGTNVGSRVYLMENDTKYQEFTLLGNEFSFDVDVSNLP
CGLNGALYFVSMDADGGVSKYPTNTAGAKYGTGYCDSQCPRDLKFINGEANVGNWTPSTNNANTGIGRY
GSCCSEMDIWEANSISNALTPHPCTTVGQSRCEADTCGGTYSSDRYAGTCDPDG
CDWNPYRLGNTSFYG
PGMTVDTTKKMTVVTQFETSGAINRFYVQDGKIIAQPNAELGSYSGNELNDDYCTAEEAEFGGSSFSDK
GGLTQFKKATSGGMVLVMSVWDDYAANMLWLDSDYPTDADPTTPGIARGTCPTTSGVPAEIEAQSPNSN
VIFSNIKFGPIGSTVPPSGGNPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGTT
CQVLNPYYSQCL
233232323133
QQACSLT
AENHPSLTWQKCTAPGSCTQQTGAVVLDANWRWVHTVSGSTNCYTGNTWDTSLCTDGKSCAQ
TCCLDGADYEGTYGVTTSGNSLKLNFVTGSNVGSRLYLMENDTKYQMFELLNREFSFDVDVSNLGCGLN
GALYFVSMDADGGMSKYSGNKAGAKYGTGYCDAQCPRDLKFINGEANVGNWTPSTNNANTGIGRYGSCC
SEMDVWEANNMATAFTPHPCTTVGQSRCEADTCGGTYSSDRYAGVCDPDGCDFNAY
RQGDKTFYGKGMT
VDTNKKMTVVTQFLTDDGTDTGTLSEIKRFYVQDGKIIANAESKIPGNPGNSITQEYCDAQEVAFGGSS
FSDKGGMAQMSKALAGPMVLVMSLWDDYAANMLWLDSDYPTDADPTTPGIARGACPTTSGVPSDVESQV
PNSYVTYSNIKFGPIGSTVPPSGGNPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCA
SGTTCQVLNPYYSQCL
333112122312 *
QQA
CTLQSETHPPLTWKRCSSGGTCSTVNGSVVLDANWRWTHTVSGSTNCYTGNQWDTSLCTDGKSCAQ
TCCLDGADYEGTYGVTTSGDSLKLNFVTQSAQKNVGSRLYLMASDTTYQEFTLLNREFTFDVDVSNLPC
GLNGALYFVSMDADGGMSKYSGNKAGAKYGTGYCDSQCPRDLKFINGEANVGNWTPSTNDANAGFGRYG
SCCSEMDVWEANSISNAVTPHPCTTVGQSRCEADTCGGTYSDNRYGGTCDPD
GCDFNPYRMGNTSFYGP
GSSFTLDTTKKLTVVTQFLTDDGTDTGTLSEIKRYYVQNGVTFQQPNSDISGVTGNSITTEFCTAQKQA
FSNTDDFNRKGGLTQFKKATSGGMVLVMSLWDDHYANMLWLDSTYPIDQAGAPGAERGTCPTTSGVPAQ
VESQSPNSYVTYSNIRFGPIGSTGNGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVC
ASGTTCQVLNPYYSQCL
10
133331223212
QQACSLTAENHPSLTWQECTSGGSCTTQNGAVVIDANWRWTHTVSGSTNCYDGNTWDTSLCTDGKSCAQ
TCCVDGAAYASTYGVTTSGSSLSIGFVTKHQYGTNVGARLYLMENDTKYQIFKLLNREFSFDVDVSQLP
CGLNGALYFVAMDADGGMSKYSGNKAGAKYGTGYCDSQCPRDLKFINGEANVGNWTPSTNNANTGIGRY
GSCCAEMDVWEANSISNAFTPHPCDTPGQSRCEADTCGGTYSSDRYAG
TCDPDGCDFNPYRQGNTSFYG
PGSSFTLDTTKKLTVVTQFLTDDGTDTGTLSEIKRYYVQNGVTFQQPNSKIPGNPGNSITQEYCDAQKV
AFGDTDDFSQHGGMAKMGAAMQQGMVLVMSLWDDYYANMLWLDSTYPTNETSSTPGAVRGTCPTTSGVP
AEIEAQSPNAKVTFSNIKFGPIGSTGNASPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGP
TVCASGTTCQVLNPYYSQCL
1311122
22121
QSACTLQSETHPPLTWQKCSSGGTCTQQTGSVVLDANWRWTHATNSSTNCYDGNTWSSTLCPDGETCAK
NCCLDGADYEGTYGVTTSGNSLKLNFVTQSAQKNVGSRLYLMASDTTYQIFKLLNREFSFDVDVSNLPC
GLNGALYFVAMDADGGVSKYPTNTAGAKYGTGYCDSQCPRDLKFINGEANVGNWTPSTNNANTGIGRYG
SCCAEMDVWEANSISEAVTPHPCDTPGQSRCEADTCGGTYSD
NRYGGTCDPDGCDFNPYRMGNTSFYGP
GKIIDTTKPFTVVTQFLTDDGTDTGTLSEIKRFYIQNSNVIPQPNSDISGVTGNSITTEFCTAQEQAFG
GSSFSDHGGLAKMGAAMQQGMVLVMSLWDDYYANMLWLDSTYPTNETSSTPGAVRGSCPTTSGVPAQVE
SQSPNSYVTYSNIKFGPINSTFTPSGGNPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPT
VCASGTTCQVLNPYYSQCL
32
2211223233
QQACTLQSETHPPLTWQECTAPGSCTTQNGSVVIDANWRWVHDVNGYTNCYTGNQWDPTYCPDDETCAQ
NCCLDGAAYASTYGVTTSGSSLSIGFVTGSNVGARLYLMASDTTYQIFKLLNREFSFDVDVSQLGCGLN
GALYFVAMDADGGVSKYPNNKAGAKYGTGYCDSQCPRDLKFIDGEANVEGWQPSSNDANAGFGDHGSCC
AEMDVWEANNMATAFTPHPCDTPGQTMCSGDDCGGTY
SDNRYGGTCDPDGCDFNAYRQGDKTFYGKGMT
VDTNKKMTVVTQFLTDDGTDTGTLSEIKRFYVQDGKIIANAESKIPGNPGNSITQEYCDAQKVAFGDTD
DFSQHGGMAKMGAAMQQGMVLVMSLWDDHYAQMLWLDSTYPIDQAGAPGAERGACPTDSGVPSDVESQV
PNAKVTFSNIKFGPIGSTVPASPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGT
TCQVLNPYYSQCL
223
311222322
QQACTLQSETHPPLTWKRCTSGGSCSTVNGSVVIDANWRWTHTVSGSTNCYTGNTWDTSLCTDDKSCAQ
TCCVDGAAYASTYGVTTSGDSLSIGFVTKHQYGTNVGARLYLMASDTTYQIFKLLNREFTFDVDVSQLP
CGLNGALYFVAMDADGGMSKYSGNKAGAKYGTGYCDSQCPRDLKFIDGEANVEGWQPSSNNANTGIGDH
GSCCAEMDVWEANSISNAVTPHPCDTPGQTMCSGDDCGGTYSDNRYGGT
CDPDGCDFNPYRMGNTSFYG
PGKIIDTTKPFTVVTQFLTDDGTDTGTLSEIKRFYIQNSNVIPQPNSDISGVTGNSITTEFCTAQKQAF
SNTDDFNRHGGLAKMGAAMQQGMVLVMSLWDDYAAQMLWLDSDYPTDADPTTPGIARGTCPTDSGVPAE
IEAQSPNAKVTFSNIRFGPINSTFTGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVC
ASGTTCQVLNPYYSQCL
12121333332
3
QSACTLQSETHPPLTWKRCTAPGSCSTVNGSVTIDANWRWVHATNSSTNCYDGNTWSSTLCPDDETCAK
NCCLDGADYSSTYGITTSGDSLNLKFVTGSNVGSRVYLMASDTTYQMFELLGNEFTFDVDVSNLGCGLN
GALYFVSMDADGGVSKYPTNTAGAKYGTGYCDAQCPRDLKFIDGEANVEGWQPSSNNANTGIGDHGSCC
SEMDVWEANNMATAFTPHPCTTVGQTMCSGDDCGGTYSDNRYGGVC
DPDGCDFNAYRQGDKTFYGKGKI
IDTNKPFTVVTQFHKNSAGVLSEIKRFYIQNSNVIPNAESKIPGNPGNSITQEYCDAQKVAFSNTDDFN
RKGGMAQMSKALAGPMVLVMSVWDDYYAQMLWLDSTYPTNETSSTPGAVRGACPTDSGVPSDVESQVPN
SNVIFSNIRFGPINSTFTGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGTTCQ
VLNPYYSQCL
11
123333232233
QQACSLTAENHPSLTWQECTSGGSCTTQNGAVTIDANWRWTHTVSGSTNCYDGNTWDTSLCTDDKSCAQ
TCCVDGADYSSTYGITTSGSSLNLKFVTKHQYGTNVGSRVYLMENDTKYQIFKLLGNEFSFDVDVSNLG
CGLNGALYFVAMDADGGMSKYSGNKAGAKYGTGYCDSQCPRDLKFIDGEANVEGWQPSSNNANTGIGDH
GSCCAEMDVWEANNMATAVTPHPCDTPGQTMCSGDDCGGTYSSDRYAGT
CDPDGCDFNAYRMGDKTFYG
KGMTVDTNKKMTVVTQFHKNSAGVLSEIKRFYVQDGKIIANAESDISGVTGNSITTEFCTAQKQAFGDT
DDFSQHGGLAKMGAAMQQGMVLVMSVWDDYYAQMLWLDSTYPTNETSSTPGAVRGACPTDSGVPAEIEA
QVPNSNVIFSNIKFGPIGSTVPASPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCAS
GTTCQVLNPYYSQCL
12
Supplementa
ry Table 2
Amino acid
sequences of 7 chimeric cellulases predicted to
be stable. T
he
H. jecorina
CBH
I
linker and cellulose binding domain are attached to
the C
-
terminus
. The chimera nomenclature is a series of numbers, each representing a
parent for the bl
ocks A
-
L. For example, chimera 123113322331 has parent 1's
sequence for block A, parent 2 for block B, etc.
222233221322
QQACSLTAENHPSLTWKRCTAPGSCSTVNGAVTIDANWRWVHDVNGYTNCYTGNTWDPTYCPDDETCAQ
NCCLDGADYSSTYGITTSGDSLNLKFVTGSNVGSRVYLMENDTKYQIFKLLNREFTFDVDVSN
LPCGLN
GALYFVAMDADGGVSKYPNNKAGAKYGTGYCDSQCPRDLKFIDGEANVEGWQPSSNNANTGIGDHGSCC
AEMDIWEANSISNALTPHPCDTPGQTMCSGDDCGGTYSSDRYAGTCDPDGCDWNPYRLGNTSFYGPGKI
IDTTKPFTVVTQFLTDDGTDTGTLSEIKRFYIQNSNVIPQPNAELGSYSGNSLNDDYCTAEKAEFSNTD
DFNRHGGLAKMGAAMQQGMVLVMSVWDDYAAQMLWLDSDYPTD
ADPTTPGIARGTCPTDSGVPSDVESQ
SPNSNVIFSNIRFGPINSTFTGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGT
TCQVLNPYYSQCL
232233222322
QQACSLTAENHPSLTWKRCTAPGSCSTVNGAVTIDANWRWVHDVNGYTNCYTGNTWDPTYCPDGETCAQ
NCCLDGADYSSTYGITTSGDSLNLKFVTGSNVGSRVYLMENDTKYQIFKLLNREFTFDVDVSNLPCGLN
GALYFVAMDADGGVSKYPNNKAGAKYGTGYCDSQCPRDLKFINGEANVGNWTPSTNNANTGIGRYGSCC
AEMDVWEANSISNAVTPHPCDTPGQSRCEADTCGGTYSSDRYAGTCDPD
GCDFNPYRMGNTSFYGPGKI
IDTTKPFTVVTQFLTDDGTDTGTLSEIKRFYIQNSNVIPQPNSDISGVTGNSITTEFCTAQKQAFSNTD
DFNRHGGLAKMGAAMQQGMVLVMSVWDDYAANMLWLDSDYPTDADPTTPGIARGTCPTTSGVPSDVESQ
SPNSNVIFSNIRFGPINSTFTGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGT
TCQVLNPYYSQCL
222211222322
QQACTLQSETHPPLTWKRCTAPGSCSTVNGSVVIDANWRWVHDVNGYTNCYTGNTWDPTYCPDDETCAQ
NCCLDGAAYASTYGVTTSGDSLSIGFVTGSNVGARLYLMASDTTYQIFKLLNREFTFDVDVSQLPCGLN
GALYFVAMDADGGVSKYPNNKAGAKYGTGYCDSQCPRDLKFIDGEANVEGWQPSSNNANTGIGDHGSCC
AEMDVWEANSISNAVTPHPCDTPGQTMCSGDDCGGTYSDNRYGGTCDPD
GCDFNPYRMGNTSFYGPGKI
IDTTKPFTVVTQFLTDDGTDTGTLSEIKRFYIQNSNVIPQPNSDISGVTGNSITTEFCTAQKQAFSNTD
DFNRHGGLAKMGAAMQQGMVLVMSLWDDYAAQMLWLDSDYPTDADPTTPGIARGTCPTDSGVPSDVESQ
SPNAKVTFSNIRFGPINSTFTGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGT
TCQVLNPYYSQCL
222223222322
QQACTATAENHPPLTWKRCTAPGSCSTVNGAVTIDANWRWVHDVNGYTNCYTGNTWDPTYCPDDETCAQ
NCCLDGADYSSTYGITSSGDSLNLKFVTGSNVGSRVYLLQDDSTYQIFKLLNREFTFDVDVSNLPCGLN
GALYFVAMDADGGVSKYPNNKAGAKYGTGYCDSQCPRDLKFIDGEANVEGWQPSSNNANTGIGDHGSCC
AEMDVWEANSISNAVTPHPCDTPGQTMCSGDDCGGTYSNDRYAGTCDPD
GCDFNPYRMGNTSFYGPGKI
IDTTKPFTVVTQFLTDDGTDTGTLSEIKRFYIQNSNVIPQPNSDISGVTGNSITTEFCTAQKQAFSNTD
DFNRHGGLAKMGAAMQQGMVLVMSVWDDYAAQMLWLDSDYPTDADPTTPGIARGTCPTDSGVPSDVESQ
SPNSNVIFSNIRFGPINSTFTGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGT
TCQVLNPYYSQCL
222211221322
QQACTLQSETHPPLTWKRCTAPGSCSTVNGSVVIDANWRWVHDVNGYTNCYTGNTWDPTYCPDDETCAQ
NCCLDGAAYASTYGVTTSGDSLSIGFVTGSNVGARLYLMASDTTYQIFKLLNREFTFDVDVSQLPCGLN
GALYFVAMDADGGVSKYPNNKAGAKYGTGYCDSQCPRDLKFIDGEANVEGWQPSSNNANTGIGDHGSCC
AEMDIWEANSISNALTPHPCDTPGQTMCSGDDCGGTYSDNRYGGTCDPD
GCDWNPYRLGNTSFYGPGKI
IDTTKPFTVVTQFLTDDGTDTGTLSEIKRFYIQNSNVIPQPNAELGSYSGNSLNDDYCTAEKAEFSNTD
13
DFNRHGGLAKMGAAMQQGMVLVMSLWDDYAAQMLWLDSDYPTDADPTTPGIARGTCPTDSGVPSDVESQ
SPNAKVTFSNIRFGPINSTFTGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGT
TCQVLNPYYSQCL
322211222322
QQACTLQSETHPPLTWKRCTAPGSCSTVN
GSVVIDANWRWVHDVNGYTNCYTGNTWDPTY
CPDDETCAQ
NCCLDGAAYASTYGVTTSGDSLSIGFVTGSNVGARLYLMASDTTYQIFKLLNREFTFDVDVSQLPCGLN
GALYFVAMDADGGVSKYPNNKAGAKYGTGYCDSQCPRDLKFIDGEANVEGWQPSSNDANAGFGDHGSCC
AEMDVWEANSISNAVTPHPCDTPGQTMCSGDDCGGTYSDNRYGGTCDPD
GCDFNPYRMGNTSFYGPGKI
IDTTKPFTVVTQFLTDDGTDTGTLSEIKRFYIQNSNVIPQPNSDISGVTGNSITTEFCTAQKQAFSNTD
DFNRHGGLAKMGAAMQQGMVLVMSLWDDHYAQMLWLDSTYPIDQAGAPGIARGTCPTDSGVPSDVESQS
PNAKVTFSNIRFGPINSTFTGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGTT
CQVLNPYYSQCL
122211222322
Q
QACTLQSETHPPLTWKRCTAPGSCSTVN
GSVVIDANWRWVHDVNGYTNCYDGNTWDPTY
CPDDETCAQ
NCCLDGAAYASTYGVTTSGDSLSIGFVTGSNVGARLYLMASDTTYQIFKLLNREFTFDVDVSQLPCGLN
GALYFVAMDADGGVSKYPNNKAGAKYGTGYCDSQCPRDLKFIDGEANVEGWQPSSNNANTGIGDHGSCC
AEMDVWEANSISNAVTPHPCDTPGQTMCSGDDCGGTYSDNRYGGTCDPDG
CDFNPYRMGNTSFYGPGKI
IDTTKPFTVVTQFLTDDGTDTGTLSEIKRFYIQNSNVIPQPNSDISGVTGNSITTEFCTAQKQAFSNTD
DFNRHGGLAKMGAAMQQGMVLVMSLWDDYYAQMLWLDSTYPTNETPTTPGIARGTCPTDSGVPSDVESQ
SPNAKVTFSNIRFGPINSTFTGPPGGNRGTTTTRRPATTTGSSPGPTQSHYGQCGGIGYSGPTVCASGT
TCQVLNPYYSQCL
14
Supplementary
Table 3.
T
A50
measurements
of
H. jecorina
CBH
I
with block G from
T. emersonii
CBH
I
and
C. thermophilum
CBH
I
.
Chimera
T
A50
(°C)
111111211111
48.3 (± 0.5)
111111311111
47.8 (± 0.1)
15
Supplementary Table 4.
Stability of
H. jecorina
CBH
I
with
the single mutation
F362M and
other
stabilizing
point
mutations.
Mutations
T
A50
(°C)
wild
-
type
46.9
(± 0.1)
F362M
49.6 (± 0.2
)
E120I, T122K
47.4 (± 0.5)
E120M, T122K
46.6 (± 0.1)
E120M, F362M
48.8 (± 0.1)
S175A, K356H
48.4 (± 0.1)
S175A, F362M
48.3
(± 0.2)
K356H, F362M
49.3 (± 0.1)
S175A, K356H, F362M
49.1 (± 0.4
)