Isolation and identification of polylactic acid degrading microorganisms from mealworm（Tenebrio molitor）gut

doi:10.3969/j.issn.1004-1524.2022.06.18

Acta Agriculturae Zhejiangensis ›› 2022, Vol. 34 ›› Issue (6): 1277-1287.DOI: 10.3969/j.issn.1004-1524.2022.06.18

• Environmental Science • Previous Articles Next Articles

Isolation and identification of polylactic acid degrading microorganisms from mealworm（Tenebrio molitor）gut

FENG Juan¹(), ZHU Tingheng²^,^*(), LUO Chunping¹, YANG Jiayue¹, ZHU Siyu¹, LI Tong¹

1. College of Agriculture and Bioengineering, Taizhou Vocational College of Science and Technology, Taizhou 318020, Zhejiang, China
2. College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, China

Received:2021-12-07 Online:2022-06-25 Published:2022-06-30
Contact: ZHU Tingheng

Abstract

Abstract:

Polylactic acid (PLA) is biodegraded rapidly under composting or thermophilic temperature but slowly under natural conditions with substantial microplastics generated. In this study, microorganisms with the ability to degrade PLA were screened from the gut of mealworm, and the degrading bacteria were identified and the degradation characteristics were determined. Mealworms was fed with PLA powder as the only food for 60 days, and the gut extract of mealworms was inoculated on the solid medium with PLA as the sole carbon source to enrich, screen and isolate PLA-degrading microorganisms. Isolated strains were identified through morphological, physiological and biochemical properties as well as ITS gene sequences analysis, the phylogenic tree was constructed to identify the taxonomic status of the strains. The screened bacteria were added to the PLA liquid fermentation medium under different nutrient conditions to determine their degradation efficiency. The results showed that 11 strains with PLA degradation potential were screened from the mealworms gut. A PLA-degrading strain (FJ001) was initially identified as the fungus Trametes hirsuta. After inoculating FJ001 in PLA thin film inorganic salt liquid medium containing 1.0% glucose for 30 days, the PLA degradation rate could reach 20.1%. In conclusion, Trametes hirsuta FJ001 was isolated from mealworms with capable of degrading PLA plastics effectively. The results suggested that strains which can degrade PLA plastics could be isolated from mealworms gut. The result showed that there were fungi that can degrade PLA in the gut of mealworm, which provided a theoretical basis for the development of microbial resources for degrading polylactic acid materials.

Key words: yellow mealworm, polylactic acid, gut microbiota, biodegradation, isolation

CLC Number:

Q93-331

FENG Juan, ZHU Tingheng, LUO Chunping, YANG Jiayue, ZHU Siyu, LI Tong. Isolation and identification of polylactic acid degrading microorganisms from mealworm（Tenebrio molitor）gut[J]. Acta Agriculturae Zhejiangensis, 2022, 34(6): 1277-1287.

Figures/Tables 7

Fig. 1 Fed on PLA or bran of mealworms A, Tenebrio molitor and its feces fed with wheat bran for 60 days; B, Tenebrio molitor fed with 350 mesh PLA film for 60 days and its feces; C, PLA film-feeding mealworm;D, Microscopic observation of feces of Tenebrio molitor fed with PLA powder at 160 times magnification;E, Microscopic observation of untreated polylactic acid powder at 160 times magnification;F, Microbial growth of Tenebrio molitor intestinal extract fed with wheat bran for 60 days inoculated in PLA inorganic salt medium; G, Growth of Tenebrio molitor intestinal extract inoculated in inorganic salt medium containing PLA for 60 days.

Fig. 2 Cutural characteristics of 10 strains on an inorganic salt medium with PLA as the only carbon source

Table 1 Cultural characteristics of the 11 strains on an inorganic salt medium with PLA as the only carbon source

菌株Strain	菌落形态Morphology		可溶性色素Soluble pigment	生长状况Growth status
LT-01	圆形/橘红Circle/Orange	无Spore-free	橘红Orange	稀少Sparse
LT-02	圆形/浓绿Circle/Dark green	浓绿孢子Dark green spore	无None	旺盛Vigorous
LT-03	不规则/橘红Orange/Wavy	无Spore-free	无None	一般Normal
LT-04	圆形/红色Circle/Pink	无Spore-free	红Red	旺盛Vigorous
YZL-01	圆形/灰Circle/Gray	灰色孢子Gray spore	无None	旺盛Vigorous
YZL-02	圆形/土黄Circle/Khaki	土黄孢子Khaki spore	无None	一般Normal
CMY-01	圆形/亮黄色Circle/Bright yellow	无Spore-free	亮黄色Bright yellow	一般Normal
CMY-02	不规则/黄Irregular/Chartreuse	黄绿孢子Chartreuse spore	黄绿Chartreuse	一般Normal
ZML-01	不规则/绿Irregular/Chartreuse	绿色孢子Green spore	绿Green	旺盛Vigorous
ZML-02	圆形/土黄Circle/Khaki	无Spore-free	无None	一般Normal
FJ001	圆形/白色Circle/White	无Spore-free	无None	旺盛Vigorous

Fig. 3 Developing conditions FJ001 strain in medium

Fig. 4 Phylogenetic tree of strain FJ001

Fig.5 Degradation performance of strain FJ001 under different nutrient conditions

Fig. 6 Scanning electron microscope images of surface change of PLA film A, PLA film control on the 10th day; B, PLA film control on the 30th day; C, PLA film degraded by FJ001 on the 10th day; D, PLA film degraded by FJ001 on the 30th day.

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Isolation and identification of polylactic acid degrading microorganisms from mealworm（Tenebrio molitor）gut

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