Sorting for secreted molecule production using a biosensor-in-microdroplet approach

Sorting large libraries of cells for improved small molecule secretion is throughput limited. Here, we combine producer/secretor cell libraries with whole-cell biosensors using a microfluidic-based screening workflow. This approach enables a mix-and-match capability using off-the-shelf biosensors through either coencapsulation or pico-injection. We demonstrate the cell type and library agnostic nature of this workflow by utilizing single-guide RNA, transposon, and ethyl-methyl sulfonate mutagenesis libraries across three distinct microbes (Escherichia coli, Saccharomyces cerevisiae, and Yarrowia lipolytica), biosensors from two organisms (E. coli and S. cerevisiae), and three products (triacetic acid lactone, naringenin, and L-DOPA) to identify targets improving production/secretion.

Candidate Gene, SmCPR1, Encoding CPR1 Related to Plant Height of the Eggplant Dwarf Mutant dwf

Eggplant is a vegetable crop with high economic value that is cultivated worldwide. The dwarf mutant is an important germplasm material that has been extensively used in crop breeding. However, no eggplant dwarf mutants have been reported, and little is known regarding the genes responsible for dwarfism in eggplant. In this study, we isolated an eggplant dwarf mutant (dwf) from an ethyl methyl sulfonate (EMS)-induced mutant library. Genetic analysis revealed that dwf was caused by a single recessive gene. A candidate gene SmCPR1, encoding cytochrome P450 reductases (CPR1), was identified by bulked segregant analysis (BSA). Mutation from G to A at 8216 bp of SmCPR1 resulted in mutation of the amino acid from valine to isoleucine. The results of KASP and Sanger sequencing further support the conclusion that SmCPR1 is a candidate gene responsible for the dwarfism of dwf. Moreover, the activity of SmCPR1 was significantly increased in dwf, which might be a response to dwarfism in dwf.

Ethyl Methyl Sulfonate-Induced Mutagenesis and Its Effects on Peanut Agronomic, Yield and Quality Traits

Peanut is an important oilseed and food crop worldwide; however, the development of new cultivars is limited by its remarkably low genetic variability. Therefore, in order to enhance peanut genetic variability, here, we treated two widely cultivated peanut genotypes, Huayu 22 and Yueyou 45, with different concentrations of the mutagen ethyl methyl sulfonate (EMS) for different durations. Based on median lethal dose (LD50) value, optimal EMS treatment concentrations for each duration were identified for each genotype. Mutants induced by EMS differed in various phenotypic traits, including plant height, number of branches, leaf characteristics, and yield and quality in plants of the M2 generation. Moreover, we identified potentially useful mutants associated with dwarfism, leaf color and shape, high oil and/or protein content, seed size and testa color, among individuals of the M2 generation. Mutations were stably inherited in M3-generation individuals. In addition to their contribution to the study and elucidation of the mechanisms underlying the regulation of the expression of some important agronomic traits, the mutants obtained in this study provide valuable germplasm resources for use in peanut improvement programs.

Catalytic synthesis of n -butyl carboxylate with immobilized ionic liquid based on response surface methodology optimization

Four kinds of functional ionic liquids (ILs) ([C 3 SO 3 Hnmp]HSO 4 ), 1-(3-sulfopropyl)-1-methylpyrrolidone phosphate ([C 3 SO 3 Hnmp]H 2 PO 4 ), 1-(3-sulfopropyl)-1-methylpyrrolidone p-toluene sulfonate ([C 3 SO 3 Hnmp]CH 3 SO 3 H) and 1-(3-sulfopropyl)-1-methylpyrrolidone methyl sulfonate ([C 3 SO 3 Hnmp]C 6 H 6 SO 3 H)) were prepared and the catalytic activity of these ILs during esterification of carboxylic acids (formic acid, acetic acid, propionic acid, butyric acid) with alcohols was investigated. The results indicated that the IL ([C 3 SO 3 Hnmp]HSO 4 ) exhibited an optimal catalytic performance. And then the IL ([C 3 SO 3 Hnmp]HSO 4 ) was immobilized to the silica gel. The immobilized IL performed more excellent catalytic activity than the unsupported [C 3 SO 3 Hnmp]HSO 4 . The effects of reaction temperature, reaction time, molar ratio of acid to alcohol and catalyst dosage were investigated. The response surface methodology based on the Box–Behnken design (BBD) was used to explore the best reaction condition of different experimental variables. Accordingly, a high n -butyl butyrate yield of 97.10% under the deduced optimal reaction conditions was obtained, in good agreement with experimental results and that predicted by the BBD model. The immobilized IL [C 3 SO 3 Hnmp]HSO 4 maintained high catalytic activity after five cycles.

Keragaan tiga jenis planlet anggrek Phalaenopsis asal Protocorm yang diinduksi Ethyl Methyl Sulfonate (EMS) secara in vitro

This research was conducted to determine the effect of concentration of Ethyl Methyl Sulfonate (EMS) to performance of three types of Phalaenopsis Planlets and to know the optimum concentration of EMS on any type of Phalaenopsis orchids. The experiment was conduc-ted at Tissue Culture Laboratory of the Faculty of Agriculture, Winaya Mukti University, Sumedang. It used a completely randomized design (CRD) with factorial pattern, that consisted of two factors and replicated twice.The first factor was the type of Phalaenopsis that resulted form hybridization which consisted of v1 (Phalaenopsis 717 X Phalae-nopsis Fire Bird), v2 (Phalaenopsis Tianong Rose X Sibling), and v3 (Phalaenopsis Luchia Pink X Phalae-nopsis Chain Xen Mammon). The second factor was the concentration of EMS that consisted of e0 (0% EMS/control), e1 (0,05% EMS), e2 (0,10% EMS), e3 (0,15% EMS), e4 (0,20% EMS), and e5 (0,25% EMS). Explant protocorm of three types of Phalae-nopsis soaked in a solution of EMS by each treat-ment for 3 hours, and cultured on MS medium Basal Modified Multiplication Shoot for 10 weeks. The experimental results showed that there are interaction between the three types of Phalae-nopsis result of a cross with a concentration of EMS to variable number of roots. Orchids P. 717 X P. Fire Bird had higher number of leaves, number of roots, leaf leang, and root length than the other. The result showed that there were interaction between three species of Phalaenopsis orchid from the crossing with EMS concentration on root variables. Orchid P. 717 X P. Fire Bird has the number of leaves, the number of roots, the number of shoots, leaf length and root length better than other types. EMS concentrations independently at all levels of treatment can not increase the number of leaves, the number of roots, fresh weight of planlet, leaf length, and root length.Keywords: Phalaenopsis, the concentration of EMS, in vitro cultureSari. Penelitian ini mempelajari dan mengetahui pengaruh konsentrasi Ethyl Methyl Sulfonate (EMS) terhadap keragaan planlet tiga jenis anggrek Phalaenopsis asal protocorm dan mencari konsentrasi optimum EMS untuk setiap jenis anggrek Phalae-nopsis secara in vitro.Percobaan dilaksanakan di Laboratorium Kultur Jaringan Fakultas Pertanian Universitas Winaya Mukti Sumedang. Eksperimen menggunakan Rancangan Acak Lengkap (RAL) Pola Faktorial, yang terdiri atas dua faktor dan diulang sebanyak dua kali.Faktor pertama adalah jenis anggrek Phalaenopsis hasil persilangan yang terdiri atas tiga jenis, yaitu v1 (Phalaenopsis 717 X Phalaenopsis Fire Bird), v2 (Phalaenopsis Tianong Rose X Sibling), dan v3 (Phalaenopsis Luchia Pink X Phalaenopsis Chain Xen Mammon). Faktor kedua adalah EMS yang terdiri atas enam taraf perlakuan, yaitu e0 (0% EMS/kontrol), e1 (0,05% EMS), e2 (0,10% EMS), e3 (0,15% EMS), e4 (0,20% EMS), dan e5 (0,25% EMS). Eksplan berupa protocorm dari tiga jenis anggrek Phalaenopsis hasil persilangan yang direndam dalam larutan EMS berdasarkan masing-masing perlakuan selama 3 jam, dan dikulturkan pada media MS Modified Multiplication Shoot Basal selama 10 minggu.Hasil penelitian menunjukkan terjadi interaksi antara tiga jenis anggrek Phalaenopsis hasil persilangan dengan konsentrasi EMS terhadap variabel jumlah akar.Anggrek P. 717 X P.Fire Bird memiliki jumlah daun, jumlah akar, jumlah tunas, panjang daun dan panjang akar yang lebih baik dibandingkan jenis lainnya. Konsentrasi EMS secara mandiri pada semua taraf perlakuan tidak dapat meningkatkan jumlah daun, jumlah akar, bobot segar planlet, panjang daun, dan panjang akar.Kata Kunci: Phalaenopsis, Konsentrasi EMS, dan Budidaya In Vitro

Supramolecular catalysis in the methylation of meta-phenylene ethynylene foldamer containing N,N-dimethylaminopyridine

DFT investigations show that the methylation reaction of N,N-dimethylaminopyridine (DMAP)-modified meta-phenylene ethynylene foldamer can be catalyzed by the noncovalent interactions between the foldamer and the methyl sulfonate esters.

Additives-Effects during Environmental Protection Methyl Sulfonic Acid Salt Bath of the Performance

Methyl sulfonate tin plating bath is a kind of high production efficiency,good environmental performance and excellent performance of the new type of tin plating bath.Research by electrochemical technology analysis an influence on electrochemical performance of the tin plating bath by adding methyl sulfonate in bath.The results show that it significantly improve the polarization properties of the bath after adding additives and the deposition potential of tin ion. The result is that polarization properties of bath perform the best when additive A is added to25mL/L,That the throwing power of the bath is 98 % and the cover ing power is 100 %.