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<table><tbody><tr><td> <h1>Table 1</h1> <table><tbody><tr><td> <p><strong>Amino acid sequences of the linker region from fungal GAs. Potential glycosylation sites in the linker sequences were predicted by the NetNGlyc and NetOGlyc programs. The predicted N-glycosylation sites are underlined and the putative O-glycosylation sites are labeled in bold.</strong></p> </td> </tr><tr><td><strong>GA</strong><br></td> <td><strong>Position (aa)</strong><br></td> <td><strong>Linker sequence</strong><br></td> </tr><tr><td></td> </tr><tr><td><em>Ro</em>GA<br></td> <td>132–167<br></td> <td>SKP<strong>TTTT</strong>A<strong>TTTTTT</strong>AP<strong>STSTTT</strong>RP<strong>SSS</strong>EPA<strong>T</strong>FP<strong>T</strong>G<u>N</u><br></td> </tr><tr><td><em>An</em>GA<br></td> <td>494–531<br></td> <td>A<strong>T</strong>GG<strong>TTTT</strong>A<strong>T</strong>P<strong>T</strong>G<strong>S</strong>G<strong>S</strong>V<strong>TSTS</strong>K<strong>TT</strong>A<strong>T</strong>A<strong>S</strong>K<strong>TSTSTSSTS</strong><br></td> </tr><tr><td><em>Hr</em>GA<br></td> <td>500–504<br></td> <td><u>N</u>VTSS<br></td> </tr><tr><td><em>Hg</em>GA<br></td> <td>490–508<br></td> <td><strong>S</strong>KQ<strong>T</strong>PNP<strong>S</strong>AAP<strong>S</strong>P<strong>S</strong>PYP<strong>T</strong>A<br></td> </tr><tr><td><em>Mc</em>GA<br></td> <td>129–177<br></td> <td>SV<strong>TTTTTT</strong>AP<strong>TTTTS</strong>GG<strong>SSTTT</strong>GG<strong>STTT</strong>A<strong>TS</strong>VP<strong>T</strong>GVPSGFP<strong>T</strong>G<u>N</u>S<strong>T</strong>ISS<br></td> </tr><tr><td><em>Nc</em>GA<br></td> <td>494–519<br></td> <td>A<strong>T</strong>A<strong>T</strong>A<strong>TS</strong>FPANL<strong>T</strong>PA<strong>STT</strong>V<strong>T</strong>PP<strong>T</strong>QTG<br></td> </tr><tr><td> <p>GenBank or GenPept accession numbers are as follows: <em>Ro</em>, <em>Rhizopus oryzae </em>(from Simpson Biotech); <em>An</em>, <em>Aspergillus niger </em>(AAP04499); <em>Hr</em>, <em>Hormoconis resinae </em>(Q03045); <em>Hg</em>, <em>Humicola grisea </em>(AAA33386); <em>Mc</em>, <em>Mucor circinelloides </em>(AAN85206); and <em>Nc</em>, <em>Neurospora crassa </em>(P14804).</p> </td> </tr><tr><td> <p>Lin <em>et al.</em><em> BMC Biochemistry</em> 2007 <strong>8</strong>:9 doi:10.1186/1471-2091-8-9</p> </td> </tr></tbody></table></td> </tr></tbody></table> <table><tbody><tr><td> <h1>Table 2</h1> <table><tbody><tr><td> <p><strong>Comparison between calculated and experimental molecular weights of GAs. The molecular masses were calculated from the deduced amino acid compositions using the ExPASy Molecular Biology Server. The experimental molecular masses were obtained from mass spectrometry or estimated from SDS-PAGE.</strong></p> </td> </tr><tr><td><strong>Protein</strong><br></td> <td><strong>Calculated mass (kDa)</strong><br></td> <td><strong>Apparent mass (kDa) (mass spectrometry/SDS-PAGE)</strong><br></td> </tr><tr><td></td> </tr><tr><td>full-length GA<br></td> <td>62.4<br></td> <td>ND/78<br></td> </tr><tr><td>T165A<br></td> <td>62.4<br></td> <td>ND/78<br></td> </tr><tr><td>N167D<br></td> <td>62.3<br></td> <td>ND/76<br></td> </tr><tr><td>GA<span>Δ</span>168–604<br></td> <td>15.2<br></td> <td>20.7/27<br></td> </tr><tr><td>GA<span>Δ</span>132–604<br></td> <td>11.7<br></td> <td>12.0/14<br></td> </tr><tr><td>GA<span>Δ</span>26–131<br></td> <td>50.7<br></td> <td>ND/69<br></td> </tr><tr><td>GA<span>Δ</span>26–145<br></td> <td>49.4<br></td> <td>59.9 to 61.1<sup>†</sup>/60<br></td> </tr><tr><td>GA<span>Δ</span>26–160<br></td> <td>47.9<br></td> <td>55.5 to 56.6<sup>‡</sup>/55<br></td> </tr><tr><td> <p>ND, not determined. <sup>†</sup>GA<span>Δ</span>26–145 displayed multiple mass spectral peaks ranging from 59.9 to 61.1 kDa. <sup>‡</sup>GA<span>Δ</span>26–160 displayed multiple mass spectral peaks ranging from 55.5 to 56.6 kDa.</p> </td> </tr><tr><td> <p>Lin <em>et al.</em><em> BMC Biochemistry</em> 2007 <strong>8</strong>:9 doi:10.1186/1471-2091-8-9</p> </td> </tr></tbody></table></td> </tr></tbody></table> <table><tbody><tr><td> <h1>Table 3</h1> <table><tbody><tr><td> <p><strong>Specific activity of wild-type <em>Ro</em>GA and truncated mutants. Specific enzyme activities were evaluated per micromole of protein.</strong></p> </td> </tr><tr><td><strong>Enzyme</strong><br></td> <td><strong>Specific activity (10</strong><sup><strong>3 </strong></sup><strong>U/<span>μ</span>mol)</strong><br></td> </tr><tr><td></td> </tr><tr><td>full-length <em>Ro</em>GA<br></td> <td>4.58 ± 0.16<br></td> </tr><tr><td>GA<span>Δ</span>26–131<br></td> <td>4.88 ± 0.04<br></td> </tr><tr><td>GA<span>Δ</span>26–145<br></td> <td>4.05 ± 0.06<br></td> </tr><tr><td>GA<span>Δ</span>26–160<br></td> <td>3.73 ± 0.33<br></td> </tr><tr><td> <p>The data are expressed as the means ± SD.</p> </td> </tr><tr><td> <p>Lin <em>et al.</em><em> BMC Biochemistry</em> 2007 <strong>8</strong>:9 doi:10.1186/1471-2091-8-9</p> </td> </tr></tbody></table></td> </tr></tbody></table> <table><tbody><tr><td> <h1>Table 4</h1> <table><tbody><tr><td> <p><strong>Oligonucleotide primers used for the construction of plasmids. Primers were used in the amplification reactions to generate various DNA fragments or mutations constructs.</strong></p> </td> </tr><tr><td><strong>Name</strong><br></td> <td><strong>Nucleotide sequence</strong><br></td> </tr><tr><td></td> </tr><tr><td>F-<em>Ro</em>GA<br></td> <td>5'-TTC<u>GAATTC</u>ATGCAATTATTCAATTTG-3'<br></td> </tr><tr><td>R-<em>Ro</em>GA<br></td> <td>5'-TTC<u>GAATTC</u>TTAAGCGGCAGGTGCACC-3'<br></td> </tr><tr><td>R-GA<span>Δ</span>168–604<br></td> <td>5'-TT<u>GAATTC</u>TCAGTTACCAGTTGGGA-3'<br></td> </tr><tr><td>R-GA<span>Δ</span>132–604<br></td> <td>5'-TT<u>GAATTC</u>TCATGTAGATACTTGGT-3'<br></td> </tr><tr><td>F-PS1<br></td> <td>5'-CGTAGTTTTTCAAGTTCTTAG-3'<br></td> </tr><tr><td>R-PS1<br></td> <td>5'-TCCTTACCTTCCAATAATTC-3'<br></td> </tr><tr><td>F-GA<span>Δ</span>132–167<br></td> <td>5'-CAAGTATCTACATCTACAATCTCC-3'<br></td> </tr><tr><td>R-GA<span>Δ</span>132–167<br></td> <td>5'-AGGAGATTGTAGATGTAGATACTTG-3'<br></td> </tr><tr><td>F-GA<span>Δ</span>26–131<br></td> <td>5'-TTGCTTGTTTCTGCTTCCAAGCCCACTACT-3'<br></td> </tr><tr><td>R-GA<span>Δ</span>26–131<br></td> <td>5'-AGTAGTGGGCTTGGAAGCAGAAACAAGCAA-3'<br></td> </tr><tr><td>F-GA<span>Δ</span>26–145<br></td> <td>5'-AGAAGTAGAAGGGGCAGCAGAAACAAGCAA-3'<br></td> </tr><tr><td>R-GA<span>Δ</span>26–145<br></td> <td>5'-TTGCTTGTTTCTGCTGCCCCTTCTACTTCT-3'<br></td> </tr><tr><td>F-GA<span>Δ</span>26–160<br></td> <td>5'-TTGCTTGTTTCTGCTGCTACTTTCCCAACT-3'<br></td> </tr><tr><td>R-GA<span>Δ</span>26–160<br></td> <td>5'-AGTTGGGAAAGTAGCAGCAGAAACAAGCAA-3'<br></td> </tr><tr><td>F-GA<span>Δ</span>26–167<br></td> <td>5'-TTGCTTGTTTCTGCTTCTACAATCTCCTCA-3'<br></td> </tr><tr><td>R-GA<span>Δ</span>26–167<br></td> <td>5'-TGAGGAGATTGTAGAAGCAGAAACAAGCAA-3'<br></td> </tr><tr><td>F-T165A<br></td> <td>5'-CCA<strong>GCT</strong>GGTAACTCTACAATCTCCTCA<br></td> </tr><tr><td>R-T165A<br></td> <td>5'-GGAGATTGTAGAGTTACC<strong>AGC</strong>TGG<br></td> </tr><tr><td>F-N167D<br></td> <td>5'-ACTGGT<strong>GAC</strong>TCTACAATCTCCTCATGGATT-3'<br></td> </tr><tr><td>R-N167D<br></td> <td>5'-GGAGATTGTAGA<strong>GTC</strong>ACCAGTTGGGAAAGT-3'<br></td> </tr><tr><td> <p>*Underlined letters indicate <em>Eco</em>RI sites. Bold letters indicate mutated nucleotides.</p> </td> </tr><tr><td> <p>Lin <em>et al.</em><em> BMC Biochemistry</em> 2007 <strong>8</strong>:9 doi:10.1186/1471-2091-8-9</p> </td> </tr></tbody></table></td> </tr></tbody></table>
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