Biotechnology1682-296x1682-2978Asian Network for Scientific Information10.3923/biotech.2008.87.93Candida tropicalis with Lyticase]]>KhorsandArezoo MaljaeiSeyed Vafa 1200871In this study, a yeast isolate was recovered from the
oral cavity. The yeast cells were converted to spheroplasts using lyticase,
instead of zymolyase, which is a cheaper and more available lytic enzyme.
Spheroplasts were subjected to metabolic lysis, mechanical disruption
and DEAE-dextran-induced-lysis. Vacuoles were then isolated by two methods
of sedimentation and flotation. Integrity and purity of isolated vacuoles
were checked by phase-contrast microscopy and determination of polyphosphate
as a vacuolar marker. Spheroplasts were found more sensitive than have
ever reported. So, they were handled very carefully. In contrast to reports
on zymolyase, spheroplast yield did not enhance with increasing length
of lytic incubation time of yeast cells in lyticase. Among the methods
used to lyse the spherplasts, mechanical disruption and treatment with
dextran were found more suitable. Despite the reports, dextran seems to
affect the released vacuoles of our strain. In addition, both isolation
methods yielded isolated vacuoles successfully and a slightly increase
in flotation method was probably due to the more spheroplasts lysis when
dextran was used or/and less vacuole disruption.]]>Bankaitis, V.A., L.M. Johnson and S.D. Emr,19868390759079Banta, L.M., J.S. Robinson, D.J. Klionsky and S.D. Emr,198810713691383Cabib, E., R. Ulane and B. Bowers,197324814511458Campbell, I. and J.H. Duffus,1988Pages: 289Pages: 289Christopher, J.R., K.R. Christopher, C.T. Yamashiro and T.H. Stevens,1991Cregg, J.M. and K.A. Russell,19981998pp: 27-39pp: 27-39Durr, M., T.H. Boller and A. Wiemken,1975105319327Ezeronye, O.U. and P.O. Okerentugba,2001Saccharomyces cerevisiae and Candida tropicalis using gut enzymes of the giant African snail Achatina achatina.]]>32190193Haas, A.,1995in vitro.]]>17283294Horst, M., E.C. Kenecht and V.P. Schu,19991028792889Kaneko, T., K. Kitamura and Y. Yamamoto,1973Arthrobacter luteus.]]>3722952302Klionsky, D.J.,19982731080710810Rothman, J.H. and T.H. Stevens,19881988Sambade, M., M. Alba, A.M. Samardon, R.W. West and M.P. Kane,200517015391551Teter, S.A., K.P. Eggerton, S.V. Scott, J. Kim, A.M. Fischer and D.J. Klionsky,200127620832087Urech, K., M. Durr, T.H. Boller, A. Wiemken and J. Schwencke,1978Saccharomyces cerevisiae.]]>116275278Walker, M.G.,19981st Edn.,pp: 62-65pp: 62-65Wang, C.W., P.E. Stromhaug, E.J. Kauffman, L.S. Weisman and D.J. Klionsky,2003163973985Weisman, L.S.,200337435460Wiemken, A. and P. Nurse,1973Candida utilis.]]>109293306Wiemken, A. and M. Durr,1974Saccharomyces cerevisiae.]]>1014557Wiemken, A., Schellenberg and K. Urech,1979Saccharomyces cerevisiae.]]>1232335Yabe, I., K.I. Houriuchi, K. Nakahara, T. Hiyama and T. Yamanaka,1999Saccharomyces cerevisiae.]]>2743490334910