Purification and characterization of cell wall lytic enzyme released by mating gametes of Chlamydomonas reinhardtii

FEBS Letters ◽  
1984 ◽  
Vol 166 (2) ◽  
pp. 293-297 ◽  
Author(s):  
Yoshihiro Matsuda ◽  
Atsuko Yamasaki ◽  
Saito Tatsuaki ◽  
Tetsuya Yamaguchi
Keyword(s):  
Cell Wall ◽  
Chlamydomonas Reinhardtii ◽  
Lytic Enzyme ◽  
Purification And Characterization

A Simple Method for Removal of the Chlamydomonas reinhardtii Cell Wall Using a Commercially Available Subtilisin (Alcalase)

2018 ◽  
Vol 28 (4) ◽  
pp. 169-178 ◽  
Author(s):  
Hyun-Ju Hwang ◽  
Yong Tae Kim ◽  
Nam Seon Kang ◽  
Jong Won Han
Keyword(s):  
Cell Wall ◽  
Chlamydomonas Reinhardtii ◽  
Algal Cell ◽  
Chemical Properties ◽  
Transformation Method ◽  
Lytic Enzyme ◽  
Higher Plant ◽  
Time Saving ◽  
Simple Method ◽  
Cell Wall Disruption

The algal cell wall is a potent barrier for delivery of transgenes for genetic engineering. Conventional methods developed for higher plant systems are often unable to penetrate or remove algal cell walls owing to their unique physical and chemical properties. Therefore, we developed a simple transformation method for <i>Chlamydomonas reinhardtii</i> using commercially available enzymes. Out of 7 enzymes screened for cell wall disruption, a commercial form of subtilisin (Alcalase) was the most effective at a low concentration (0.3 Anson units/mL). The efficiency was comparable to that of gamete lytic enzyme, a protease commonly used for the genetic transformation of <i>C. reinhardtii</i>. The transformation efficiency of our noninvasive method was similar to that of previous methods using autolysin as a cell wall-degrading enzyme in conjunction with glass bead transformation. Subtilisin showed approximately 35% sequence identity with sporangin, a hatching enzyme of <i>C. reinhardtii</i>, and shared conserved active domains, which may explain the effective cell wall degradation. Our trans­formation method using commercial subtilisin is more reliable and time saving than the conventional method using autolysin released from gametes for cell wall lysis.

scholarly journals Purification and Characterization of Soluble and Cell Wall-bound Acid .ALPHA.-Glucosidases of Ripe Yellow Banana Pulp.

2001 ◽  
Vol 48 (1) ◽  
pp. 19-25
Author(s):  
Yotaro Konishi ◽  
Mia Harada ◽  
Mie Nakasuji ◽  
Marisa D'Innocenzo ◽  
Franco M. Lajolo
Keyword(s):  
Cell Wall ◽  
Purification And Characterization

Purification and Characterization of Extracellular and Cell Wall Bound β-Glucosidases fromAspergillus kawachii

1998 ◽  
Vol 62 (10) ◽  
pp. 1938-1946 ◽  
Author(s):  
Kazuhiro IWASHITA ◽  
Kouichi TODOROKI ◽  
Hitoshi KIMURA ◽  
Hitoshi SHIMOI ◽  
Kiyoshi ITO
Keyword(s):  
Cell Wall ◽  
Purification And Characterization

Purification and characterization of a .gamma.-like DNA polymerase from Chlamydomonas reinhardtii

Biochemistry ◽  
1991 ◽  
Vol 30 (4) ◽  
pp. 1127-1131 ◽  
Author(s):  
Z. F. Wang ◽  
Junming Yang ◽  
Z. Q. Nie ◽  
Madeline Wu
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Dna Polymerase ◽  
Purification And Characterization
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