Research Objective
1. Mutation breeding of industrial microorganisms
Use ion beam, plasma and other physical methods, breed high-yield industrial microbial strains. Research the ion beam and plasma mutagenesis mechanism.
2. Fermentation and metabolic control
Use of principles and methods of metabolic engineering, molecular biology and reaction engineering, optimize the culture conditions and Increase the yield and production capacity of industrial bio-products.
3. Microbial (enzymatic) catalysis
Microorganisms (enzyme) catalyzed complex substrate and modified the molecule structure, can produce high value-added products. Research the mechanism in the interaction between Microorganisms (enzyme) and the substrate.
4. Biological active product separation and purification
Study the pretreatment of the fermentation broth, the technical problems in the process of extraction, refining, product processing. Research the highly selective separation and purification of active products of microbial fermentation technology and reduce the cost of separation and purification.

Research Progress
1. Achievements in industrial strains
By Ion beam implantation modified technique, Vitamin C and Arachidonic Acid have been applied into industrialization successfully and has substantial beneficial effects. For years our studies, antibacterial lipopeptide production strain Bacillus subtilis (activity increased and fermentation time decreased, Appl. Microbiol. Biotechnol. 2005); surfactin high yield strains (production increased three times, Plasma Sci. Technol. 2006); L-lactic acid high yield strains (resistance high temperature strains, J. Microbiol. Biotechnol. 2004；resistance acid strains, Plasma Sci. Technol. 2007；utilize xylose strain, Plasma Sci. Technol. 2007); chitosanase high yield strains(Ind. Microbiol. Biotechnol. 2006；L. Appl. Microb. 2008).

2. Research of Ubiquinone-10 biosynthesis pathway modification by Metabolic Flux Analysis
As an evaluation standard of screening model and breeding tactic based on the MFA, the characteristics of mutants were examined, including genetic stability, change of intracellular metabolic flux distribution, cellar activity and profiles of fermentation, in the last part of this paper. The results indicated that the screening model and breeding strategy were goodness and reasonable. At the same time, the ion beam irradiation breeding technique, which introduced metabolic engineering principles and methods, was discussed. We believe that ion beam irradiation breeding technique would become more effective by employing the metabolic engineering. On the other hand, the former would not only extend the researching fields but provide a new investigation way for the latter. Metabolic flux analysis of Agrobacterium tumefaciens ATCC 4452 and ATX12 are studied in order to better understand the metabolic pathway of CoQ10. The data are shown: G-6-P and pyruvic acid are two improtant nodes in the production of CoQ10. The flux from G-6-P to Ru5P in ATX12 is twice that of ATCC4452, and the flux from PYR to DDP is not increasing greatly when the total flux of PYR decreases. The content of CoQ10 is increased with the rise of Biomass by ion beam implantation. In the metabolic network, the enzyme for producing DDP may be the key point for CoQ10 production. （Appl. Microbiol. Biotechnol., 2006; Process Biochemistry, 2006；Plasma Science and Technology, 2008）

 
3. Studies of Lipopeptide Antibiotics Produced by Bacillus subtilis JA
Bio-pesticide is an important alternative of chemical pesticides according to the sustainable development of agriculture. Bacillus subtilis JA is a biocontrol candidate isolated by our lab which shows antagonistic activity against a broad spectrum of plant pathogens. In this study, JA was tested for its ability to antagonize the growth of various plant and post-harvest pathogens in vitro. The antibiotics produced by JA were isolated and characterized. The volatile compounds emitted from JA were analyzed and the way the volatiles antagonizing the growth of pathogenic fungus was studied. The antagonistic effects of volatiles generated by B. subtilis JA were investigated. Both spore germination and elongation of germ tubes in B. cinerea were significantly inhibited by volatiles in vitro. The volatiles caused protoplasm retraction in B. cinerea from the hyphal tips to the spores. The volatile compounds were analyzed by Gas chromatography/mass spectrometry (GC/MS). Fourteen comounds were identified. The results showed that the volatiles generated by B. subtilis JA were rich in antifungal compounds. ( Biotechnol Lett, 2008)

4. The solution of foaming problems in biosurfactants fermentation
The serious foam is one of important factor to inhibit the normal microbial fermentation process. It becomes more serious in these high yield and efficient biosurfactants production, such as surfactin. Foams are created by the rapid stirring and aerating in aerobic fermentations, and are stabilised by the presence of surfactants in the culture broth. Foaming creates problems including the undesirable stripping of product, nutrients, and cells into the foam, and can make process containment difficult. Foam suppression is commonly achieved by the addition of chemical antifoams, but it can reduce the oxygen transfer efficiency, and may exert adverse effects on the cell physiology, and can be costly. By analyzing the characteristics of the biosurfactant fermentation, mechanism of foam initiation, and shortcoming of traditional bioreactor, a modified bioreactor with two foam collectors and product separation system was designed to deal with fermentation foam problem and end-product feedback inhibition. The results indicated that the modified bioreactor got an advantage over the normal one by increasing significantly the surfactin concentration and shortening the fermentation period. The design of two collectors and recycling by gas pressure can resolve the serious foam problem and was suitable for large-scale fermentation and general application. By manipulating the gas pressure of the two collectors, the process of liquefying of foam was enhanced. Foam and liquid broth in the second collector were transmitted into bioreactor rapidly by air pressure. （Food Technol. Biotechnol. 2009）

 
5. Research of chitosanase and glucanase
(1)Chitin, the β-(1-4)-linked homopolymer of N-acetyl-D-glucosamine, is the second most abundant polysaccharide which is only next to that of cellulose. Chitosan, a D-glucosamine polymer, is totally or partially deacetylated derivative of chitin. Chitosan can be converted to watersoluble oligosaccharides by chemical or enzyme. With its advantages in environmental compatibility, low cost and reproducibility, chitosanase hydrolysis becomes more and more popular in recent years. However, the utility of chitosanase in such hydrolysis is limited because of its cost and unavailability in bulk quantity. For the production of oligosaccharides from chitosan, a chitosanase-producing ray fungi was isolated from soil. On the basis of phylogenetic analysis and 18SrDNA gene sequence, it was identified as Gongronella JG. It is the only one Gongronella strain that can degrade chitosan reported so far. Chitosanase was homogeneously purified by CM- sepharose FF fast flow anion exchange , Sephacryl S200 followed by SP-Sepharose FF, and the molecular weight was 96kDa according to SDS-PAGE. Enzyme analysis showed that the optimum pH and temperature of Gongronella JG. （J. Agric. Food Chem. 2006）（L. Appl. Microb. 2008）

(2)Endo-β-l, 4-glucanase is one of the most important cellulases, and it has been widely used in various industrial purposes such as in the paper, textile and brewing industries. The stability and substrate specificity of cellulase are of great importance for its industrial application. Typically, cellulase from bacterial and fungal consists of a cellulose-binding domain and a catalytic domain. And these two domains are separated by a long and flexible peptide sequence and they can perform their functions independently. CBDs endow the enzymes with high affinity for native, ordered cellulose, while the catalytic domain performs hydrolysis of substrates. Removal of CBD affects cellulase activity, thermal and pH stability to different extents. Lots of cellulases from Bacillus species have been studied and evaluated for biotechnological applications. Here we focused on the effect of cellulase binding domain(CBD) and linker on characterization of an endo-β-l, 4-glucanase from Bacillus subtilis strain JA18. Compared with Egl22-499, the characterization of Egl22-330, including the optimal reaction temperature and pH, doesn’t change. They also show the similar pH stability. But the thermal stability of Egl22-330 is much better than Egl22-499. （Bioresource Technology 2009）

 
6、Research of lactic acid
L(+)-lactic acid is a typical organic acid widely applied in food, chemical and pharmaceutical industries. Meanwhile, it can also be used to synthesize polylactic acid (PLA), which is thought to be used in manufacturing new biodegradable plastic. However, commercial replacing plastic with PLA depends on whether L(+)-LA can be produced at a lower cost than petrochemical derivatives. Mutagenic breeding was taken with the method of ions implanting. In the case of mixed substrates, when xylose and glucose were present in a bubble column, L(+)-lactic acid conversion rate reached 80%.

 
Production of L-lactic acid from fermentation broth by bipolar membrane electrodialysis (EDBM) with three-compartment was proposed and investigated. We studied the effects of operation voltage, flow rate and initial concentration of L-sodium lactate on some parameters such as conversion ratio of sodium lactate, loss rate, energy consumption and current efficiency in the processes. The results showed that the final L-lactic acid concentration of 144.31g/L was reached when fed-batch electrodialysis was performed to product L-lactic acid from the broth containing100.25 g/L of L-sodium lactate under the optimum conditions of operation voltage at 15V and flow rate at 40L/h. The conversion ratio of sodium lactate, loss rate, energy consumption and current efficiency were 81.22%, 1.5%, 0.81kwh/kg and 91.8%，respectively. Electrodialysis wastewater supplemented with glucose could be reused as a fermentation medium for L-lactic acid production in the bioreactor.（Plasma Science＆Technology 2008，J. Ind. Microb. Biotech., 2009） 

7、The study on microbial diesel and its derived fine chemicals
Biodiesel production using straw as raw material, is a potential path to resolve the limitation of raw materials on microbial oil fermentation. Through several rounds of mutagenesis by nitrogen ion beam implantation, we have obtained a Mortierella alpina (named as Mortierella alpina I502-8, MAI502-8) which can efficiently utilize xylose as carbon source for the production of microbial oil. The mutant have some interesting characteristics, for example, it can cumulate fat by utilizing the mixed sugar (glucose/xylose=5/3, w/w) as carbon source. Both biomass and cell fat content in the mutant were reach to 29.8 g/L and 11.7 g/L through high density fermentation and metabolic regulation following raw sugar utilization rate of 99.4%. Compared to original strain, biomass and fat content increased to 2.59 times and 2.05 times respectively. We have also primarily researched on the techniques of microbial oil refining and fractionation. It was finded that 70 percent biodiesel and 30 percent of the high-end oils were obtained by wet fractionation, high-end oils’ recovery rate was reach to 80-90%. Prepared biodiesel quality was in line with the quality standards of the National Biodiesel BD100 and American ASTM D6751-2003 standard. This research provided a new channel for the bio-energy conversion from straw waste. Meanwhile, the results have important theoretical and practical significance for both comprehensive utilization of straw waste and bio-diesel technology development (Preparative Biochmistry & Biotechnology, 2007; Chemical, Engineering & Technology, 2011).

8. Biocatalytic synthesis of propyl gallate by Transesterification in Organic medium
Biocatalysis in anhydrous medium is an important subdivision of industrial biocatalysis. Presently, to improve biocatalytic efficiency of enzyme is one of key challenges in biocatalytic technology. Propyl gallate (PG) is an excellent antioxidant and is also considered as important pharmaceutical intermediates.

In view of the flexibility of enzymatic conformation in aqueous phase and its rigidity in organic medium, imprinting technique is competent to hyper-activate enzyme in anhydrous phase. Imprinting, cryogenic protection, and immobilization techniques were complexly applied to improve the transesterification-catalyzing performance of tannase in organic media. The results showed that such treatments as pH tunning, substrate imprinting, and interfacial activation all hyper activate tannase and improve its biocatalytic performance by 123 folds relative to the original control. Furthermore, immobilizing the imprinted tannase remarkably relieves its aggregation caused by the imprinting, and thus the immobilization increases the apparent activity of the imprinted enzyme and makes its conversion rate of substrate (CR) reach 40 %. On the other hand, combinational application of Triton X-100, mannose, and magnesium ions reduces the damage of enzyme activity resulted from the lyophilization in the protocol of imprinting and promotes its CR up to 49%. Not only does the study broaden the application fields of imprinting technique but also presents a reference for modification of enzyme in the future.

The analysis on thermostability of the imprinted tannase found that its activation energy of irreversible thermal inactivation (Ed) and half-time are 85.54 kJ mol−1 and 1710 h, respectively, which are more than the previous reports. The thermodynamic analysis of the imprinted tannase indicated that the free energy of Gibbs (ΔG) and enthalpy (ΔH) in enzyme-catalyzing transesterification at 40-60 ◦C are 97.1~98.4 kJ mol−1 and 82.77~82.94 kJ mol−1, respectively, which are lower than the free energy stack from hydrolysis and esterification both catalyzed by tannase. This reveals the biocatalytic efficiency of transesterification (direct synthesis) is higher than that of the two-step biocatalysis composed of hydrolysis and esterification. Entropy of activation of denaturation (ΔS) in the direct synthesis is -0.047~-0.045 kJ mol−1 K-1 at 40-60 ◦C. It indicates that the reaction cannot run spontaneously. The kinetic analysis of the imprinted tannase showed the Km at 40 ◦C is 0.054 mM, which is less than that in tannic acid-hydrolyzing reaction by tannase. It is inferred that the imprinting enhances the biocatalytic performance of enzyme by improving its affinity to substrate.

The strengthened measures as imprinting and cryogenic protection not only is widespreadly suitable to biocatalysis in organic media, but also gives a experience means to biosynthesize a few chemicals in organic media.

 
9. Research of filamentous fungi morphology metabolite engineering
Filamentous microorganisms are the most common microorganisms used for antibiotic production on an industrial scale. As a filamentous fungus, P. chrysogenum always grows by hyphal extension and branching, two processes for which regulation is still not completely understood. Previous studies have shown that penicillin secretion is correlated with hyphal extension rates and tip growth and, as such, morphological characterization of P. chysogenum is of interest.

 
Chitin synthases catalyze the formation of β-(1,4)-glycosidic bonds between N-acetylglucosamine residues to form the unbranched polysaccharide chitin, which is the major component of cell walls in most filamentous fungi. Several studies have shown that chitin synthases are structurally and functionally divergent and play crucial roles in the growth and morphogenesis of the genus Aspergillus although little research on this topic has been done in Penicillium chrysogenum. Several studies have shown that chitin synthases are structurally and functionally divergent and play crucial roles in the growth and morphogenesis of the genus Aspergillus although little research on this topic has been done in Penicillium chrysogenum. We used BLAST to find the genes encoding chitin synthases in P. chrysogenum related to chitin synthase genes in Aspergillus nidulans. Three homologous sequences coding for a class III chitin synthase CHS4 and two hypothetical proteins in P. chrysogenum were found. The gene whose product showed the highest identity and encoded the class III chitin synthase CHS4 was studied in detail. To investigate the role of CHS4 in P. chrysogenum morphogenesis, we developed an RNA interference system to silence the class III chitin synthase gene chs4. After transformation, mutants exhibited a slow growth rate and shorter and more branched hyphae, which were distinct from those of the original strain. The results also showed that the conidiation efficiency of all transformants was reduced sharply and indicated that chs4 is essential in conidia development. The morphologies of all transformants and the original strain in penicillin production were investigated by light microscopy, which showed that changes in chs4 expression led to a completely different morphology during fermentation and eventually caused distinct penicillin yields, especially in the transformants PcRNAi1-17 and PcRNAi2-1 where penicillin production rose by 27% and 41%, respectively.

Research Programs
1. Vitamin K2 industrial fermentation optimization, The National High Technology Research and Development Program ("863"Program 2014AA021704)
2 Devolepment of tannase and its application in synthesis of propyl gallate by transesterification. Knowledge Innovation Project of The Chinese Academy of Sciencesthe (KSCX2-YW-G-050)
3.Antibiotic fermentation morphological information and control systems，The National High Technology Research and Development Program ("863"Program 2009AA02Z305)
4. Research of vitamin K2 biosynthesis pathway modification by Low-energy Ion Beam Irradiation Based on Metabolic Flux Analysis（Y09FCQ5121）
5. (KSCX2-YW-G-050)
6. Optimize Jinggangmycin microbial fermentation and the products purification
7. Research of kinetics in Morphology of filamentous microorganisms in industrial fermentation（2008B002）
8. Ion beam modification and metabolic regulation in high-yield strains of microorganisms diesel (HY-DLZ/JC-2008YZJJ-3)
9. Steam-exploded straw two-step fermentation of high-purity L-lactic acid（07010202076）
10. Pilot and industrialization of new biosurfactants
11. Ion beam implantation L-lactic acid producing strain selection and fermentation（2006AA020102）
12. Research of Ubiquinone-10 biosynthesis pathway modification by Low-energy Ion Beam Irradiation Based on Metabolic Flux Analysis (20576132)

Achievements and Publication
1. Genhai Zhao, Jun Dai, Peng Wang, Guohong Gong, Li Wang, Hui Liu, Zhiming Zheng*. An efficient method for the enrichment of the arachidonicacid methyl ester from Mortierella alpina-derived crude oils. Food Bioprod Process, 91:617-622, 2013
2. Genhai Zhao, Guohong Gong, Peng Wang, Li Wang, Hui Liu, Zhiming Zheng*, Enzymatic synthesis of L-aspartic acid by Escherichia coli cultured with a cost-effective corn plasm medium. Ann Microbiol, doi: 10.1007/s13213-014- 0805-3, 2014
3. Liu, Zhiming Zheng*, Peng Wang, Guohong Gong, Li Wang, Genhai Zhao, Morphological changes induced by class III chitin synthase gene silencing could enhance penicillin production of Penicillium chrysogenum, Appl Microbiol Biotechnol, 97:3363-3372, 2013
4. Hui Liu, Peng Wang, Guohong Gong, Li Wang, Genhai Zhao, Zhiming Zheng*, Morphology engineering of Penicillium chrysogenum by RNA silencing of chitin synthase gene, Biotechnol Lett, 35(3), 423-429, 2013
5. Hui Liu, Peng Wang, Yihua Hu, Guohong Gong, Genhai Zhao, Junying Song, Jun Dai, Zhiming Zheng*. Construction of an RNAi expression vector and transformation into Penicillium chrysogenum. Ann Microbiol, 64:113-120, 2014
6. Peng Wang, Zhen Chen, Juan Li, Li Wang, Guohong Gong, Genhai Zhao, Hui Liu, Zhiming Zheng*, Immobilization of Rhizopus oryzae in a modified polyvinyl alcohol gel for L(+)-lactic acid production, Ann Microbiol, 63:957-964, 2013
7. Yan Liu, Zhenglian Xue, Zhiming Zheng*, Guohong Gong, Peng Wang, Guangjun Nie, He-Ne laser irradiation of folate-producing Candida utilis and optimization of culture conditions under submerged fermentation, Ann Microbiol, 63(2): 641-647, 2013.
8. Guangjun Nie, Xiaoran Yang, Hui Liu, Li Wang, Guohong Gong, Wei Jin, Zhiming Zheng*, N+ Ion beam implantation of tannase-producing Aspergillus niger and optimization of its process parameters under submerged fermentation, Ann Microbiol, 63(1), 279–287, 2013
9. Guangjun Nie, Hui Liu, Zhen Chen, Peng Wang, Genhai Zhao, Zhiming Zheng*, Synthesis of propyl gallate from tannic acid catalyzed by tannase from Aspergillus oryzae: Process optimization of transesterification in anhydrous media, J Mol Catal B Enzym, 82, 102-108, 2012
10. Guangjun Nie, Zhiming Zheng*, Guohong Gong, Genhai Zhao, Yan Liu, Junying Song, Jun Dai, Characterization of bioimprinted tannase and its kinetic and thermodynamics properties in synthesis of propyl gallate by transesterification in anhydrous medium, Appl Biochem Biotechnol, 167(8), 2305-2317, 2012
11. Guangjun Nie, Zhiming Zheng*, Wei Jin, Guohong Gong, Li Wang, Development of a tannase biocatalyst based on bio-imprinting for the production of propyl gallate by transesterification in organic media, J Mol Catal B Enzym, 78, 32-37, 2012
12. Jin Wei, Nie guangjun, Liu Hui, Yang Xiaoran, Gong Guohong, Wang Li, Zhiming Zheng*, Improving Aspergillus niger tannase yield by N+ ion beam implantation, Braz Arch Biol Technol, 56(1): 135-142, 2013
13. Gui Fang, Wang Hui, Wang Peng, Liu Hui, Cai Xiaochun, Hu Yihua, Yuan Chengling, Zheng Zhiming*, The mutation breeding and mutagenic effect of air plasma on Penicillium Chrysogenum, Plasma Sci Technol, 14(4), 61-66, 2012
14. Wang Peng, Zhang Lamei, Zheng Zhiming*, Wang Li, Wang Hui, Yuan Chengling, Gong Guohong, The co-fermentation study of glucose and xylose for microbial lipid production with Mortierella alpina by low-energy ion beam implantation, Chem Eng Technol, 34( 3), 422-428, 2011
15. Wang Peng, Xiaoni Zhang, Li wang, Zhang zhen, Mingli Tang , Jiabin Li. Subinhibitory concentrations of ciprofloxacin induce SOS response and mutations of antibiotic resistance in bacteria, Ann Microbiol, 2010, 60:511–517
16. Wang Peng, Li Juan, Wang Li, Tang Mingli, Yu Zengliang, Zheng Zhiming. L(+)-Lactic acid production by co-fermentation of glucose and xylose with Rhizopus oryzae obtained by low-energy ion beam irradiation, Journal of Industrial Microbiology & Biotechnology, 2009, 36(11): 1363-1368.
17. Yujuan Wang, Hang Yuan, Jun Wang, Zengliang Yu, Truncation of the Cellulose Binding Domain Improved Thermal Stability of Endo-β-1, 4-glucanase From Bacillus subtilis JA18 ；Bioresource Technology, 2009, 100: 345-349.
18. G.H. Gong, Z.M. Zheng, H. Chen, C.L. Yuan, P. Wang, L.M. Yao, Z.L. Yu. Enhanced production of surfactin by Bacillus subtilis E8 mutant obtained by ion beam implantation. Food Technol. Biotechnol. 2009, 47(1): 27-31.

19. Hua Chen,Xiang Xiao,Jun Wang,Lijun Wu ,Zhiming Zheng ,Zengliang Yu, Antagonistic effects of volatiles generated by Bacillus subtilis on spore germination and hyphal growth of the plant pathogen, Botrytis cinerea.Biotechnol Lett, 2008, 30: 919-923.
20. XU Dejun, ZHENG Zhiming, WANG Peng,WANG Li, YUAN Hang, YU Zengliang, Breeding of Coenzyme Q10 Produced Strain by Low-Energy Ion Implantation and Optimization of Coenzyme Q10 Fermentation.Plasma Science and Technology, 2008,10(6): 758-763.
21. H. Chen, L. Wang, C.X. Su, G.H. Gong, P. Wang and Z.L.Yu, Isolation and characterization of lipopeptide antibiotics produced by Bacillus subtilis.Letters in Applied Microbiology, 2008, 47: 180-186.
22. Fan Yonghong，Yang Yingge， Zheng Zhiming .Enhancement of L(+)-Lactic Acid Production of Immobilized Rhizopus oryzae Implanted by Ion Beam. Plasma Science＆Technology, 2008, 10: 115-119.
23. W. Zhou, H. Yuan, J. Wang and J. Yao Production, purification and characterization of chitosanase produced by Gongronella sp. JG Letters in Applied Microbiology. 2008. 46(1): 49-54.
24. LI Wen, Zheng Zhiming, Yu Zengliang. Optimization of L(+)-lactic Acid Fermentation without Neutralisation of Rhizopus Oryzae Mutant RK02 by Low-Energy Ion Implantation. Plasma Science & Technology, 2008, 10(2): 260-264.
25. YANG Yingge, FAN Yonghong, LI Wen, WANG Dongmei,WU Yuejin, ZHENG Zhiming,YU Zengliang, Optimization of L(+)-Lactic Acid Production from Xylose with Rhizopus Oryzae Mutant RLC41-6 Breeding by Low-Energy Ion Implantation. Plasma Science and Technology, 2007, 9(5): 638-642.
26. Chengling Yuan,Xiangqin Wang,and Zengliang Yu, Separation and Preparative Purification of Arachidonic Acid from Microbial Lipids by Urea Inclusion Reaction and HPLC. Preparative Biochmistry & Biotechnology, 2007,37: 149-159.
27. Shao-Bin Gu, Jian-Ming Yao, Qi-Peng Yuan, Pei-Jian Xue, Zhi-Ming Zheng, Li Wang, Zeng-Liang Yu, A novel approach for improving the productivity of ubiquinone-10 producing strain by low-energy ion beam irradiation. Appl Microbiol Biotechnol, 2006, 72: 456-461。
28. Shao-Bin Gu, Jian-Ming Yao, Qi-Peng Yuan, Pei-Jian Xue, Zhi-Ming Zheng, Zeng-Liang Yu, Kinetics of Agrobacteriumtumefaciens ubiquinone10batch production. Process Biochemistry, 2006, 41:1908-1912.
29. Caixin Su, Wei Zhou, Yonghong Fan, Li Wang,Shiguang Zhao, Zengliang Yu, Mutation breeding of chitosanase-producing strain Bacillus sp. S65 by low-energy ion implantation. J. Ind Microbiol Biotechnol, 2006, 33:1037-1042.
30. Caixin Su, Dongmei Wang, Liming Yao And Zengliang Yu, Purification, Characterization, and Gene Cloning of a Chitosanase from Bacillus Species Strain S65. J. Agric.Food Chem, 2006, 54: 4208-4214.
31. Liu Qingmei, Yuan Hang, Wang Jun, Gong Guohong, Zhou Wei. A Mutant of Bacillus subtilis with High-producing Surfactin by Ion Beam Implantation. Plasma Science & Technology. 2006, 8: 491–496.
32. Liu J., Liu M., Wang J., Yao J.M., Pan R.R., Yu Z.L. Enhancement of the Gibberella zeae growth inhibitory lipopeptides from a Bacillus subtilis mutant by ion beam implantation. Appl. Microbiol. Biotechnol. 2005, 69(2): 223-238.
33. Liu Qingmei, Yao Jianming, Pan Renrui, Yu Zengliang. A mutant strain of a surfactant-producing bacterium with increased emulsification activity. Plasma Science & Technology. 2005, 7(3): 2885-2892.
34. Liu J., Yao J.M. Study on mutagenic breeding of Bacillus subtilis and properties of its antifungal substances. Plasma Science & Technology. 2004, 6(4): 2433-2436.
35. Yuan Hang, Zhou Wei, Wang Jun, Liu Qingmei, Zhang Shuqing etc. Enhancement of Gongronella sp. JG Chitosanase production by Ion Beam Implantation. Plasma Science & Technology. 2007, 9(1): 115-118.
36. Chunmei Ge, Shaobin Gu, Xiuhong ZHOU et al. Breeding of L(+)-Lactic Acid Producing Strain by Low-Energy Ion Implantation. J. Microbiol. Biotechnol. 2004, 14(2): 363-366.

Patent and software copyrights
1. patent: A method of breeding Penicillium chrysogenum morphological mutant strains (201310132497.8)
2. patent: A method for changing the yield and morphology of fungal spores (201310162551.3)
3. patent: A method for efficient production of vitamin K2 by Flavobacterium (201310280268.0)
4. patent: A processing technology for polyunsaturated fatty acids produced from microorganisms (CN102504962A)
5. patent: A processing technology for Rhizopus oryzae immobilized in a modified polyvinyl alcohol gel for L(+)-lactic acid production (ZL200910116455.9)
6. patent: A foam back fermentation system (200920187963.1).
7. patent: Effectively reduce avermitilis fermentation cycle to improve the production of avermectin (200610086279.5)
8. patent: A magnetic polymer flocculation pretreatment of L-lactic acid fermentation broth (200610041238.4)
9. patent: Bacillus subtilis antimicrobial peptide separation and purification methods (200410014483.7)
10. software copyrights: The mycelium microscopy image segmentation and feature extraction software (2011R11S044664)