By Michael E. Himmel, John O. Baker, John N. Saddler
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Gene constitution of a bifunctional cellulase gene (celA) remoted from Teredinobacter turnerae / S.N. Freer, R.V. Greene, and R.J. Bothast --
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Additional resources for Glycosyl Hydrolases for Biomass Conversion
C Units are micromoles of glucose reducing sugar equivalents formed from CMC per minute at 25°C d In the presence of cellulose, residual activity has been observed after a 30 minute exposure to 100°C(/7). ; ACS Symposium Series; American Chemical Society: Washington, DC, 2000. ch003 and agar (15 g). 01 g/ml of Ostrazin Brilliant Red-hydroxyethyl Cellulose (OBRC). Positive clones were selected based upon the formation of clear halos that surrounded the colonies. Molecular Biology Procedures. Standard methods were used for DNA isolation, subcloning, and restriction enzyme analysis (19).
9746-9751. The activities of the mutant enzymes on CMC, SC, filter paper (FP) and 2,4dinitrophenyl cellobioside (DNPCB) are shown in Table III. Clearly Aspll7 functions as the catalytic acid since all three mutant enzymes have very low activity on CMC and SC, but near wild-type activity on DNPCB which has a good leaving group. These results show that the mutants are able to carry out every function normally except protonation of the leaving group. The Asp 156 Ala and Asn mutant enzymes also have near wild-type activity on DNPCB, but somewhat reduced activity on CMC and SC, showing that Asp 156 only affects the catalytic acid.
ACS Symposium Series; American Chemical Society: Washington, DC, 2000. ch003 --iLFMFWYDLPNRDCAALASNGELRISEDGFNIYKSDYIAPIVEl ^ K A ^ p V P L ^ ^ 150 Cat 160 fimi LL Τ fusca gMwjcJF, IAAJlEÎbsLPNlflT^îlsEllA Cat 1Ë§ 230 -QAAPY 240 250 260 STTAyFASgDIGFV^DVjA G P 290 Cat M- 270 300 310 320 330 192 ΙβΑ 189 280 262 vgGl§N§LiI§^§KWVD 258 340 350 2 [WNSYLAEKPFVTOWRSAMISKGMPSSIGMLIDTA C fimi ~]PS S I GMI§Dlj^GWœpS|RP'l}s ι τΜ TDV|IS^Y]\|DA fusca | F R ¥ 3 I GMLI DTSRNGWGGHNRpJGpSdaTE 360 370 380 390 400 410 420 -IPGIDAYVWVKPQGESDGVSDPNFEIDPNDPNKQHDPMCDH-FASNSS^ fimi Τ fusca |p_gGYAASHL D A § V W J | K E P G E S D G J |A 430 Tt Cat C fimi Τ fusca 255 S^TINNl|lfï[RS^FYE| Τ C 185 IDJ 2 |AHSGMLGWSDNFAQGVNLIYEVVANLGSGINPIAGFVSNSANYTPVEEPFLPDANLQVGGQPVRSSDFYE| :SGWLGV}DsbËGPSAl|lJFA Tt 210 | C Q E A N G P G G Y R D G I R H A I T E L G K I PNVYSYVD3J !
Glycosyl Hydrolases for Biomass Conversion by Michael E. Himmel, John O. Baker, John N. Saddler