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Phyllosphere yeasts rapidly break down biodegradable plastics.

Kitamoto HK, Shinozaki Y, Cao XH, Morita T, Konishi M, Tago K, Kajiwara H, Koitabashi M, Yoshida S, Watanabe T, Sameshima-Yamashita Y, Nakajima-Kambe T, Tsushima S - AMB Express (2011)

Bottom Line: The use of biodegradable plastics can reduce the accumulation of environmentally persistent plastic wastes.Strains of P. antarctica isolated from leaves and husks of paddy rice displayed strong degradation activity on these films at 30°C.Reliable source of biodegradable plastic-degrading microorganisms are now in our hands.

View Article: PubMed Central - HTML - PubMed

Affiliation: National Institute for Agro-Environmental Sciences (NIAES), 3-1-3 Kannondai, Tsukuba, Ibaraki 305-8604 Japan. kitamoto@affrc.go.jp.

ABSTRACT
The use of biodegradable plastics can reduce the accumulation of environmentally persistent plastic wastes. The rate of degradation of biodegradable plastics depends on environmental conditions and is highly variable. Techniques for achieving more consistent degradation are needed. However, only a few microorganisms involved in the degradation process have been isolated so far from the environment. Here, we show that Pseudozyma spp. yeasts, which are common in the phyllosphere and are easily isolated from plant surfaces, displayed strong degradation activity on films made from poly-butylene succinate or poly-butylene succinate-co-adipate. Strains of P. antarctica isolated from leaves and husks of paddy rice displayed strong degradation activity on these films at 30°C. The type strain, P. antarctica JCM 10317, and Pseudozyma spp. strains from phyllosphere secreted a biodegradable plastic-degrading enzyme with a molecular mass of about 22 kDa. Reliable source of biodegradable plastic-degrading microorganisms are now in our hands.

No MeSH data available.


Related in: MedlinePlus

SDS-PAGE of biodegradable plastic-degrading enzyme (PaE) from Pseudozyma antarctica JCM 10317 and a commercially available lipase, CALB-L. (A) Purification of PaE. M Molecular mass standards; 1 Ammoniun sulfate precipitate; Fractions of 2 passed through the DEAE-Sepharose column; 3 eluated from SP-Sepharose column; 4 Gel filtration. SDS-PAGE was performed using 12% gel and detected by silver staining (B) Purified PaE and CALB-L. M Molecular mass standards; 1 purified PaE (0.25 μg); 2 CALB-L (25 μg). SDS-PAGE was performed using 14.1% gel and detected by CBB. The black arrow in the PaE lane shows the PaE isolated from the culture medium. The black arrow in the CALB-L lane indicates lipase B. The white arrows indicate impurities.
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Figure 5: SDS-PAGE of biodegradable plastic-degrading enzyme (PaE) from Pseudozyma antarctica JCM 10317 and a commercially available lipase, CALB-L. (A) Purification of PaE. M Molecular mass standards; 1 Ammoniun sulfate precipitate; Fractions of 2 passed through the DEAE-Sepharose column; 3 eluated from SP-Sepharose column; 4 Gel filtration. SDS-PAGE was performed using 12% gel and detected by silver staining (B) Purified PaE and CALB-L. M Molecular mass standards; 1 purified PaE (0.25 μg); 2 CALB-L (25 μg). SDS-PAGE was performed using 14.1% gel and detected by CBB. The black arrow in the PaE lane shows the PaE isolated from the culture medium. The black arrow in the CALB-L lane indicates lipase B. The white arrows indicate impurities.

Mentions: The strain P. antarctica JCM10317 is synonymous with Candida antarctica CBS 5955, and known to produce two lipases, lipase A (43 kDa) and lipase B (33 kDa), on culture media containing oil (Ishii 1988). As the culture conditions required for producing lipases and PaE were identical, we postulated that these common lipases from the strain could degrade biodegradable plastic. However, from a culture medium of P. antarctica JCM10317, we purified an enzyme that degraded biodegradable plastic (PaE) and had a molecular weight of about 22 kDa (Table 3, Figure 5). At pH 6.8, the relative degradation activity of commercially available lipase B (CALB-L) on PBSA emulsion was about 1/6500th that of PaE. We confirmed by MS fingerprinting of PaE and lipase B that PaE was a different protein from lipases A and B (data not shown). Esterase activites of PaE against pNP-butyrate and pNP-palmitate are 715.33 ± 124.44 U/mg PaE and 194.72 ± 36.11 U/mg PaE, respectively, showing that PaE is an esterase with wide spectrum to degrade esterified bond of long-chain fatty acids.


Phyllosphere yeasts rapidly break down biodegradable plastics.

Kitamoto HK, Shinozaki Y, Cao XH, Morita T, Konishi M, Tago K, Kajiwara H, Koitabashi M, Yoshida S, Watanabe T, Sameshima-Yamashita Y, Nakajima-Kambe T, Tsushima S - AMB Express (2011)

SDS-PAGE of biodegradable plastic-degrading enzyme (PaE) from Pseudozyma antarctica JCM 10317 and a commercially available lipase, CALB-L. (A) Purification of PaE. M Molecular mass standards; 1 Ammoniun sulfate precipitate; Fractions of 2 passed through the DEAE-Sepharose column; 3 eluated from SP-Sepharose column; 4 Gel filtration. SDS-PAGE was performed using 12% gel and detected by silver staining (B) Purified PaE and CALB-L. M Molecular mass standards; 1 purified PaE (0.25 μg); 2 CALB-L (25 μg). SDS-PAGE was performed using 14.1% gel and detected by CBB. The black arrow in the PaE lane shows the PaE isolated from the culture medium. The black arrow in the CALB-L lane indicates lipase B. The white arrows indicate impurities.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3293741&req=5

Figure 5: SDS-PAGE of biodegradable plastic-degrading enzyme (PaE) from Pseudozyma antarctica JCM 10317 and a commercially available lipase, CALB-L. (A) Purification of PaE. M Molecular mass standards; 1 Ammoniun sulfate precipitate; Fractions of 2 passed through the DEAE-Sepharose column; 3 eluated from SP-Sepharose column; 4 Gel filtration. SDS-PAGE was performed using 12% gel and detected by silver staining (B) Purified PaE and CALB-L. M Molecular mass standards; 1 purified PaE (0.25 μg); 2 CALB-L (25 μg). SDS-PAGE was performed using 14.1% gel and detected by CBB. The black arrow in the PaE lane shows the PaE isolated from the culture medium. The black arrow in the CALB-L lane indicates lipase B. The white arrows indicate impurities.
Mentions: The strain P. antarctica JCM10317 is synonymous with Candida antarctica CBS 5955, and known to produce two lipases, lipase A (43 kDa) and lipase B (33 kDa), on culture media containing oil (Ishii 1988). As the culture conditions required for producing lipases and PaE were identical, we postulated that these common lipases from the strain could degrade biodegradable plastic. However, from a culture medium of P. antarctica JCM10317, we purified an enzyme that degraded biodegradable plastic (PaE) and had a molecular weight of about 22 kDa (Table 3, Figure 5). At pH 6.8, the relative degradation activity of commercially available lipase B (CALB-L) on PBSA emulsion was about 1/6500th that of PaE. We confirmed by MS fingerprinting of PaE and lipase B that PaE was a different protein from lipases A and B (data not shown). Esterase activites of PaE against pNP-butyrate and pNP-palmitate are 715.33 ± 124.44 U/mg PaE and 194.72 ± 36.11 U/mg PaE, respectively, showing that PaE is an esterase with wide spectrum to degrade esterified bond of long-chain fatty acids.

Bottom Line: The use of biodegradable plastics can reduce the accumulation of environmentally persistent plastic wastes.Strains of P. antarctica isolated from leaves and husks of paddy rice displayed strong degradation activity on these films at 30°C.Reliable source of biodegradable plastic-degrading microorganisms are now in our hands.

View Article: PubMed Central - HTML - PubMed

Affiliation: National Institute for Agro-Environmental Sciences (NIAES), 3-1-3 Kannondai, Tsukuba, Ibaraki 305-8604 Japan. kitamoto@affrc.go.jp.

ABSTRACT
The use of biodegradable plastics can reduce the accumulation of environmentally persistent plastic wastes. The rate of degradation of biodegradable plastics depends on environmental conditions and is highly variable. Techniques for achieving more consistent degradation are needed. However, only a few microorganisms involved in the degradation process have been isolated so far from the environment. Here, we show that Pseudozyma spp. yeasts, which are common in the phyllosphere and are easily isolated from plant surfaces, displayed strong degradation activity on films made from poly-butylene succinate or poly-butylene succinate-co-adipate. Strains of P. antarctica isolated from leaves and husks of paddy rice displayed strong degradation activity on these films at 30°C. The type strain, P. antarctica JCM 10317, and Pseudozyma spp. strains from phyllosphere secreted a biodegradable plastic-degrading enzyme with a molecular mass of about 22 kDa. Reliable source of biodegradable plastic-degrading microorganisms are now in our hands.

No MeSH data available.


Related in: MedlinePlus