Biochemical characterization of chloromethane emission from the wood- rotting fungus Phellinus pomaceus

Deepti Saxena, Saleh Aouad, Jihad Attieh, Hargurdeep S. Saini

Research output: Contribution to journalArticle

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Abstract

Many wood-rotting fungi, including Phellinus pomaceus, produce chloromethane (CH3Cl). P. pomaceus can be cultured in undisturbed glucose mycological peptone liquid medium to produce high amounts of CH3Cl. The biosynthesis of CH3Cl is catalyzed by a methyl chloride transferase (MCT), which appears to be membrane bound. The enzyme is labile upon removal from its natural location and upon storage at low temperature in its bound state. Various detergents failed to solubilize the enzyme in active form, and hence it was characterized by using a membrane fraction. The enzyme had a sharp pH optimum between 7 and 7.2. Its apparent K(m) for Cl- (ca. 300 mM) was much higher than that for I- (250 μM) or Br- (11 mM). A comparison of these K(m) values to the relative in vivo methylation rates for different halides suggests that the real K(m) for Cl- may be much lower, but the calculated value is high because the CH3Cl produced is used immediately in a coupled reaction. Among various methyl donors tested, S-adenosyl-L-methionine (SAM) was the only one that supported significant methylation by MCT. The reaction was inhibited by S-adenosyl-L-homocysteine, an inhibitor of SAM-dependent methylation, suggesting that SAM is the natural methyl donor. These findings advance our comprehension of a poorly understood metabolic sector at the origin of biogenic emissions of halomethanes, which play an important role in atmospheric chemistry.

Original languageEnglish
Pages (from-to)2831-2835
Number of pages5
JournalApplied and Environmental Microbiology
Volume64
Issue number8
Publication statusPublished - 1 Aug 1998
Externally publishedYes

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Phellinus pomaceus
methyl chloride
Methyl Chloride
S-Adenosylmethionine
S-adenosylmethionine
methylation
Methylation
Fungi
fungus
enzyme
transferases
fungi
Enzymes
enzymes
membrane
atmospheric chemistry
halides
biogenic emission
Peptones
Membranes

Cite this

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title = "Biochemical characterization of chloromethane emission from the wood- rotting fungus Phellinus pomaceus",
abstract = "Many wood-rotting fungi, including Phellinus pomaceus, produce chloromethane (CH3Cl). P. pomaceus can be cultured in undisturbed glucose mycological peptone liquid medium to produce high amounts of CH3Cl. The biosynthesis of CH3Cl is catalyzed by a methyl chloride transferase (MCT), which appears to be membrane bound. The enzyme is labile upon removal from its natural location and upon storage at low temperature in its bound state. Various detergents failed to solubilize the enzyme in active form, and hence it was characterized by using a membrane fraction. The enzyme had a sharp pH optimum between 7 and 7.2. Its apparent K(m) for Cl- (ca. 300 mM) was much higher than that for I- (250 μM) or Br- (11 mM). A comparison of these K(m) values to the relative in vivo methylation rates for different halides suggests that the real K(m) for Cl- may be much lower, but the calculated value is high because the CH3Cl produced is used immediately in a coupled reaction. Among various methyl donors tested, S-adenosyl-L-methionine (SAM) was the only one that supported significant methylation by MCT. The reaction was inhibited by S-adenosyl-L-homocysteine, an inhibitor of SAM-dependent methylation, suggesting that SAM is the natural methyl donor. These findings advance our comprehension of a poorly understood metabolic sector at the origin of biogenic emissions of halomethanes, which play an important role in atmospheric chemistry.",
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Biochemical characterization of chloromethane emission from the wood- rotting fungus Phellinus pomaceus. / Saxena, Deepti; Aouad, Saleh; Attieh, Jihad; Saini, Hargurdeep S.

In: Applied and Environmental Microbiology, Vol. 64, No. 8, 01.08.1998, p. 2831-2835.

Research output: Contribution to journalArticle

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AU - Aouad, Saleh

AU - Attieh, Jihad

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AB - Many wood-rotting fungi, including Phellinus pomaceus, produce chloromethane (CH3Cl). P. pomaceus can be cultured in undisturbed glucose mycological peptone liquid medium to produce high amounts of CH3Cl. The biosynthesis of CH3Cl is catalyzed by a methyl chloride transferase (MCT), which appears to be membrane bound. The enzyme is labile upon removal from its natural location and upon storage at low temperature in its bound state. Various detergents failed to solubilize the enzyme in active form, and hence it was characterized by using a membrane fraction. The enzyme had a sharp pH optimum between 7 and 7.2. Its apparent K(m) for Cl- (ca. 300 mM) was much higher than that for I- (250 μM) or Br- (11 mM). A comparison of these K(m) values to the relative in vivo methylation rates for different halides suggests that the real K(m) for Cl- may be much lower, but the calculated value is high because the CH3Cl produced is used immediately in a coupled reaction. Among various methyl donors tested, S-adenosyl-L-methionine (SAM) was the only one that supported significant methylation by MCT. The reaction was inhibited by S-adenosyl-L-homocysteine, an inhibitor of SAM-dependent methylation, suggesting that SAM is the natural methyl donor. These findings advance our comprehension of a poorly understood metabolic sector at the origin of biogenic emissions of halomethanes, which play an important role in atmospheric chemistry.

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