Susceptibilities of Yeasts to Yeast Cell Wall Lytic Enzyme of<i>Arthrobacter luteus</i>

Kaneko, Tatsuhiko; Kitamura, Kumpei; Yamämoto, Yasushi

doi:10.1080/00021369.1973.10861005

Cited by 11 publications

(12 citation statements)

References 3 publications

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“…Mercaptoethanol was required for the action of Zymolyase on living cells of Candida lipolytica; cysteine, sodium thioglycollate and sodium sulphide enhanced lysis to a lesser extent, whereas dithiothreitol, L-ascorbic acid and cysteamine were ineffective (Kaneko et al, 1973). When young mycelia of Geotrichum candidum were treated with mercaptoethanol prior to the digestion by a lytic enzyme complex of Streptomyces satsumaensis, cell wall lysis was markedly accelerated (Dooijewaard-Kloosterziel et al, 1973).…”

Section: Discussionmentioning

confidence: 99%

Formation and Regeneration of Protoplasts from Conidiobolus lamprauges

Ishikawa

Ōishi

1985

Microbiology

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Protoplasts were formed from aged germlings and mycelia of Conidiobolus lurnpruuges, using a combination of chitinase and fl-glucanase. Methanol, ethanol and propanol stimulated protoplast formation, apparently by a mechanism different from that of mercaptoethanol. Regeneration of protoplasts in soft agar was detectable by light microscopy 2 h after incubation and colonies were observable by the naked eye within 40 h; the regeneration rate was 65%.

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Section: Discussionmentioning

confidence: 99%

Formation and Regeneration of Protoplasts from Conidiobolus lamprauges

Ishikawa

Ōishi

1985

Microbiology

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“…The effects of yeast cell wall-digesting enzymes on P. pastoris are not well understood, and yeast species can vary widely in their sensitivity to yeast lytic enzyme (24). Furthermore, Saccharomyces cerevisiae spheroplasts prepared by lyticase treatment exhibited a resistance to lysis that was dependent on growth conditions (2,24) and could show an increase in optical absorbance during lysis (26). Consequently, in order to reduce variability in yeast lytic enzyme sensitivity and protoplasting efficiency, the same strain of P. pastoris and the same growth medium were used in all experiments.…”

Section: Resultsmentioning

confidence: 99%

In Vivo Functional Assay of a Recombinant Aquaporin inPichia pastoris

Daniels

Wood

Yeager

2006

Appl Environ Microbiol

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The water channel protein PvTIP3;1 (␣-TIP) is a member of the major intrinsic protein (MIP) membrane channel family. We overexpressed this eukaryotic aquaporin in the methylotrophic yeast Pichia pastoris, and immunogold labeling of cellular cryosections showed that the protein accumulated in the plasma membrane, as well as vacuolar and other intracellular membranes. We then developed an in vivo functional assay for water channel activity that measures the change in optical absorbance of spheroplasts following an osmotic shock. Spheroplasts of wild-type P. pastoris displayed a linear relationship between absorbance and osmotic shock level. However, spheroplasts of P. pastoris expressing PvTIP3;1 showed a break in this linear relationship corresponding to hypo-osmotically induced lysis. It is the difference between control and transformed spheroplasts under conditions of hypo-osmotic shock that forms the basis of our aquaporin activity assay. The aquaporin inhibitor mercury chloride blocked water channel activity but had no effect on wild-type yeast. Osmotically shocked yeast cells were affected only slightly by expression of the Escherichia coli glycerol channel GlpF, which belongs to the MIP family but is a weak water channel. The important role that aquaporins play in human physiology has led to a growing interest in their potential as drug targets for treatment of hypertension and congestive heart failure, as well as other fluid overload states. The simplicity of this assay that is specific for water channel activity should enable rapid screening for compounds that modulate water channel activity.The first water channel gene was cloned serendipitously during the characterization of human blood group antigens and was found to encode a putative channel in the red blood cell plasma membrane. When the protein was expressed in Xenopus laevis oocytes, its function as a water channel was suggested by a concomitant increase in the swelling rate of osmotically shocked cells (41). This demonstration of water channel activity ended decades of speculation that proteins were responsible for the high water permeability observed in certain biological membranes.Over the last decade many organisms have been shown to possess a class of protein channels, termed "aquaporins," which are specialized to facilitate the transcellular movement of water. Aquaporins are members of the major intrinsic protein (MIP) superfamily found in all organisms, from archaebacteria to animals (27). The family progenitor is thought to have arisen from the tandem duplication of a three-transmembrane spanning domain protein with an Asn-Pro-Ala consensus sequence duplicated between the two halves of the protein (45). Generally, aquaporins facilitate the movement of water across cell membranes in response to osmotic gradients, functioning in cellular and organismal osmoregulation and solute transport (5, 32).MIP family proteins are small (ϳ28 kDa) and are usually found as homotetramers. Recent high-resolution structures determined by electron crystallogra...

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“…Segundo Kaneko et al (1973), as células de leveduras são mais susceptíveis à lise enzimática quando tratadas com -mercaptoetanol ou cisteína. A fração A da preparação comercial Zymolyase, composta por uma -1,3 glucanase, é capaz de causar lise da parede celular de leveduras somente na presença demercaptoetanol.…”

Section: Efeito Da Cisteína E -Mercaptoetanol Na Lise De Levedura Comunclassified

β-1,3 Glucanases e quitinases: aplicação na lise de leveduras e inibição de fungos

Fleuri

Sato

2008

Ciênc. agrotec.

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RESUMOObjetivou-se, no presente trabalho, a aplicação de -1,3 glucanases e quitinases da linhagem Cellulosimicrobium cellulans 191 na lise de leveduras e inibição de fungos, respectivamente. O delineamento experimental mostrou que as melhores condições para a lise de Saccharomyces cerevisiae KL-88 pela -1,3 glucanase foi pH 6,5 e 35ºC. As células de leveduras incubadas por 10 h em frascos sem agitação mostraram-se mais susceptíveis à lise pela ação da enzima. Foi obtido maior lise da levedura quando a suspensão de células foi submetida ao tratamento com -1,3 glucanase e cisteína 1mM. A enzima invertase intracelular ou ligada à célula de S. cerevisiae KL-88 e K. marxianus NCYC 587 foi extraída após tratamento da suspensão celular com -1,3 glucanase, sendo que o tratamento prévio das leveduras com a enzima aumentou a susceptibilidade das células à lise com ultra-som. A preparação de quitinase foi capaz de formar halos de inibição de alguns fungos.Termos para indexação: -1,3 glucanases, quitinases, lise celular, inibição. ABSTRACTThe aim of this work was the application of -1,3 glucanases and chitinases by Cellulosimicrobium cellulans 191 strain on yeast cell lysis and fungi inhibition, respectively. The experimental design study showed that the best conditions to Saccharomyces cerevisiae KL-88 lysis by -1,3 glucanase extract were pH 6,5 and 35ºC. This study also demonstrated that the yeast cells were more susceptible to lysis after 10 h of cultivation in flasks without agitation. Lysis activity was increased when S. cerevisiae KL-88 cell suspension was treated with -1,3 glucanase and cystein 1mM. The enzyme invertase of S. cerevisiae KL-88 and Kluyveromyces marxianus NCYC 587 was extracted after treatment of cell suspension with -1,3 glucanase and the previous treatment of yeasts with the enzyme, increased the susceptibility to lysis when ultrasonic treatment was used. The chitinase presented growth inhibition halos for some of the fungi.

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Susceptibilities of Yeasts to Yeast Cell Wall Lytic Enzyme ofArthrobacter luteus

Cited by 11 publications

References 3 publications

Formation and Regeneration of Protoplasts from Conidiobolus lamprauges

Formation and Regeneration of Protoplasts from Conidiobolus lamprauges

In Vivo Functional Assay of a Recombinant Aquaporin inPichia pastoris

β-1,3 Glucanases e quitinases: aplicação na lise de leveduras e inibição de fungos

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